RU2299887C2 - Methods for synthesis of aplidine and novel anti-tumor derivatives, methods for their industrial preparing and using - Google Patents

Abstract

FIELD: organic chemistry, antibiotics, medicine, oncology.

SUBSTANCE: invention relates to novel derivatives of aplidine possessing anti-tumor activity and represented by the general formula:

EFFECT: valuable medicinal property of compound, improved methods of synthesis and preparing.

28 cl, 3 tbl, 121 ex

Description

The present invention relates to methods for the synthesis of aplidine and new antitumor derivatives, methods for their industrial production and use.

Aplidine has the following cyclic structure with a side chain:

Didemnins form a class of cyclic depsipeptides that have been isolated from various species of Trididemnum genus (Rinehart, Jr. et al. J, Am. Chem. Soc. 103, 1857-59 (1981), Rinehart, Jr. et al. Science, 212, 933-935 (1981) and have antitumor and antiviral activities, of which aplidine is one of the most active natural antideancer didemnins, and USP 5.834.586 describes the isolation and antitumor activity of aplidine.

A number of synthetic and natural analogues of aplidine have been described (Rinehart, Jr. et al. J. Med. Chem, 1996, 39, 2819-2834), including various modifications of the side chain while maintaining the same macrocycle structure.

A recently described related structure of didemnins, called tamandarins (Fenical, W. et al, J, Org. Chem., 2000, 65, 782-792), isolated from unidentified ascidium compounds of the didemnid family. It was found that these molecules differ only in the presence of hydroxyisovalerianic acid (Hiv ³ ), instead of hydroxyisovalerylpropionic acid (Hip ³ ). These compounds are described as highly active antiviral, antitumor and immunosuppressive peptides.

The present invention relates to the above compounds, called derivatives of aplidine, intended for use in medicine, in particular for the treatment of tumors. The invention also relates to pharmaceutical preparations comprising the compounds intended for the treatment of tumors, for example, solid tumors, and the use of the compounds in the manufacture of a medicament for the treatment of tumors. Particularly preferred is the treatment of solid tumors, such as tumors of the bladder, breast, colon or colon, stomach, liver, nscl = nmr (non-small cell lung cancer), ovary, pancreas, pharynx, prostate, kidney, scl = mrl ( small cell lung cancer), retinoblastoma, melanoma, fibrosarcoma, chondrosarcoma or osteosarcoma, or treatment of leukemia / lymphomas such as ALL (promyelocytic leukemia), ALL (acute lymphoblastic), CML (chronic myelogenous), ALL (B-cell), leukemia ( capillary b-cell g), leukemia (plasma cell), lymphoma (T-cell), lymphoma (T-skin cells), lymphoma (undifferentiated), lymphoma (Burkitt B cells), lymphoma (histiocytic), lymphoma (B-cell), lymphoma ( ascites burkitt).

Examples of pharmaceutical compositions of the invention include any solid (e.g., tablets, pills, capsules, granules) or liquid forms (solutions, suspensions or emulsions) with a suitable composition suitable for oral, local or parenteral administration, and these compositions may contain a pure compound or in combination with any carrier or other pharmacologically active compounds. When administered parenterally, these compositions must be sterile.

For convenience, the compound may be conjugated to a carrier protein or other suitable vehicle for delivery to an animal or human body. Conjugation can be carried out directly between the carrier and the compound or indirectly, through a suitable coupling agent.

Administration of a compound or compositions of the present invention can be carried out by any suitable method, such as intravenous infusion, oral administration of drugs, intraperitoneal or intravenous administration. Applicants prefer to use an infusion time of up to 24 hours, more preferably 2-12 hours, and most preferably 2-6 hours. A short infusion time allowing treatment without leaving the hospital overnight is especially desirable. However, if necessary, the infusion can last 12-24 hours or even longer. The infusion can be performed at suitable intervals, say in 2-4 weeks. Pharmaceutical compositions containing a compound of the invention can be delivered by liposomal or nanosphere encapsulation, in sustained release formulations, or other standard delivery methods.

The exact dose of the compound varies depending on the specific composition, method of application, specific location, recipient, and requiring treatment of a carcinoma or tumor. Other factors, such as age, body weight, gender, diet, time of administration, rate of excretion, recipient condition, drug combinations, sensitivity reactions, and disease severity should also be taken into account. The introduction can be carried out continuously or periodically within the maximum permissible dose.

The compounds of this invention can be used with other drugs to provide combination therapy. Additional drugs may be part of the same composition, or provided as a separate composition for administration at the same time or at another time. The identity of the complementary drug is not specifically limited, and suitable examples include:

a) drugs with anti-mitotic effects, especially those that target cytoskeletal elements, including microtubule modulators, such as taxane drugs (such as taxol, paclitaxel, taxotere, docetaxel), podophyllotoxins or vinca alkaloids (vincristine, vinblastine);

b) antimetabolic drugs such as 5-fluorouracil, cytarabine, gemcitabine, purine analogs such as pentostatin, methotrexate);

c) alkylating agents such as mustard nitrogen compounds (such as cyclophosphamide or ifosfamide);

d) DNA targeting drugs, such as anthracycline drugs: adriamycin, doxorubicin, farmorubicin or epirubicin;

e) drugs targeting topoisomerase, such as etoposide;

f) hormones and hormone agonists or antagonists such as estrogens, antiestrogens (tamoxifen and related compounds) and androgens, flutamide, leuprorelin, goserelin, cyprotron or octreotide;

g) drugs aimed at signal transduction in tumor cells, including antibody derivatives such as herceptin;

h) alkylating drugs, such as platinum-based drugs (cis-platinum, carbonplatinum, oxaliplatin, paraplatin) or nitrosoureas;

i) drugs potentially affecting tumor metastases, such as matrix metalloproteinase inhibitors;

j) gene therapy and antisensory agents;

k) antibody based therapeutics; and

l) other biologically active compounds of marine origin, especially ecteinascidins, such as ET-743.

One aspect of the present invention relates to compounds of the formula:

Where:

X independently means —C (R) ₂ -, —O—, —S— or —NR—, where R independently means H or an organic group selected from an alkyl group, an alkenyl group, an aryl group, an aralkyl group and substituted derivatives thereof, which are substituted by one or more heterocyclic group, alkoxy group, hydroxy group, mercapto group, optionally protected amino group, guanidino group or halogen group;

X ₂ independently means CR, O (and R _{2 is} absent), S (and R _{2 is} absent) or N, where R independently means H or an organic group selected from an alkyl group, an alkenyl group, an aryl group, an aralkyl group and substituted derivatives thereof which are substituted by one or more heterocyclic group, alkoxy group, hydroxy group, mercapto group, optionally protected by an amino group, guanidino group or a halogen group;

Y is - (COR ′) _n CO—, where n is 0 or 1 and R ′ is an organic group selected from an alkyl group, an alkenyl group, an aryl group, an aralkyl group and substituted derivatives thereof, which are substituted with one or more heterocyclic groups, an alkoxy group, a hydroxy group, a mercapto group, an optionally protected amino group, a guanidino group or a halogen group;

X ₁ means O or S;

each of R ₁ , R ₂ and R ₄ independently means H or an organic group selected from an amido group of RCONH - or an acyl group of RCO -, where R means, as indicated, an alkyl group, an alkenyl group, an aryl group, an aralkyl group and substituted derivatives, which are substituted by one or more heterocyclic group, alkoxy group, hydroxy group, mercapto group, optionally protected amino group, guanidino group or halogen group, and R ₁ or R ₂ when X ₂ means N, and R ₄ may additionally mean -SO ₂ R, where R accepts above signs cheniya;

or R ₁ and R ₂ can form optionally N-substituted proline with X ₂ , N-substituted proline aa8 has the formula

where R ₃ independently means H or an organic group selected from the group RSO ₂ - or an acyl group RCO-, where R is as defined herein, or R ₃ is an alkyl group, an alkenyl group, an aryl group, an aralkyl group and substituted derivatives that are substituted with one or more a carbonyl group, a hydroxy group, a mercapto group, an optionally protected amino group, a guanidino group or a halogen group;

or R ₁ and R ₂ may form, with X _{2, a} cycloalkyl, aryl or heterocyclic group optionally substituted with one or more R ₃ groups;

or R ₁ , R ₂ , X ₂ , R _4, and nitrogen carrying R ₄ can form an oxadiazaspiroalkane N-substituted with an R ₅ group, where R ₅ independently is H or an organic group selected from the RSO ₂ - or RCO acyl group -, where R means, as indicated, an alkyl group, an aryl group, an aralkyl group and substituted derivatives which are substituted by one or more carbonyl group, alkoxy group, hydroxy group, mercapto group, amino group, guanidino group or halogen group;

or aa8 is a substituted organic group selected from the group RSO ₂ - or an acyl group RCO-, where R is, as indicated, an alkyl group, an aryl group, an aralkyl group and substituted derivatives which are substituted by one or more carbonyl group, alkoxy group, hydroxy group, a mercapto group, amino group, guanidino group or halogen group;

and their pharmaceutically acceptable salts.

Preferred compounds include those in which X is —NR—, where R is as defined. More preferably, X is —NH— or —NMe— and most preferably X is —NH—.

Other preferred compounds include those in which X is —O—.

The Y group preferably means —COR′CO—, where R ₁ is an alkyl group, in particular where R ′ is —CHCH ₃ -.

Other preferred compounds include those compounds in which Y is —CO—.

In view of these preferences, a preferred class of compounds is that class where X is —NH— or —O— and Y is —CHCHCH ₃ CO— or —CO—.

Preferably R ₄ is methyl.

Preferably X ₁ means = O.

Preferably, X ₂ R ₁ is an optionally substituted aralkyloxy group, such as a benzyloxy group.

Other preferred compounds include those compounds in which X ₂ R ₁ is an optionally substituted amino group, more preferably those in which X ₂ R ₁ is an —NHR ₁ group, where R ₁ is an optionally substituted alkyl group, alkenyl group, aryl group, or an aralkyl group, in particular an alkyl group or an aryl group, such as a phenyl group or a butyl group.

Additional preferred compounds include those compounds in which X ₂ R ₁ means an optionally substituted alkyl group, in particular where X ₂ R ₁ means a propyl group, an isopropyl group, a pentyl group or a biotin group.

A group of preferred compounds are those compounds in which —C (= X ₂ ) R ₁ R ₂ forms an optionally substituted acyl group of an amino acid. A suitable optionally substituted acyl group of an amino acid is optionally substituted proline or optionally substituted glycine, or optionally substituted valine, and more specifically, optionally substituted proline means optionally substituted norvalineproline, optionally substituted alanineproline, Boc-proline, optionally substituted alkylproline, or optionally substituted glycine means heterocyclic substituted glycine, or optionally substituted valine, means valine, Boc-valine or alkyl valine. Preferably, the optionally substituted proline means norvaline proline, Z-norvaline proline, alanine proline, Z-alanine proline, Boc-alanine proline, isobutyryl proline or optionally protected D-lactylproline, or heterocyclic substituted glycine means coumarin glycine, or optionally substituted valine valine or valine valine.

Another group of preferred compounds includes those compounds in which X ₁ is S and X ₂ R ₁ is an —NHR ₁ group, where R ₁ is an optionally substituted alkyl group, alkenyl group, aryl group or an aralkyl group. R ₁ preferably means an alkyl group or an aryl group, more preferably a phenyl group or a butyl group.

R ₁ and R ₂ can form a heterocyclic group with X ₂ , optionally substituted with one or more R ₃ groups. For example, a heterocyclic group may be coumarin.

Preferred compounds include those in which aa8 is substituted with an organic group RSO ₂ -, where R is as defined, such as methyl.

R ₁ , R ₂ , X ₂ , R ₄ and nitrogen bearing R ₄ can form an oxadiazaspiroalkane, N-substituted by R ₅ , where R ₅ is H. N-substituted oxadiazaspiroalkane preferably means 6-oxa-1,7-diazaspiro [4, 4] nonan.

Examples of compounds of the present invention include:

3- [Aip] -Z-didemnin A,

8- [phenylurea] didemnin A,

8- [butylurea] didemnin A,

3- [val] -8- [isobutyryl] aplidine,

9- [norvaline] aplidine,

3- [Hiv] -9- [isobutyryl] aplidine,

3- [Val] -9- [isobutyryl] aplidine,

3- [hiv] -8- [isobutyryl] didemnin A,

3- [Hiv] -9- [Ala] aplidine,

3- [Hiv] -9- [Nva] aplidine,

8- [phenylthiourea] didemnin A,

8- [coumarin] didemnin A,

8- [butylthiourea] didemnin A,

3- [Hiv] -9- [D-Lac] aplidine,

8- [methylsulfonyl] didemnin A,

3- [val] -Z-didemnin A,

3- [Hiv] -8- [Val] didemnin A,

3- [Hiv] -8- [butyryl] aplidine,

3- [val] didemnin A,

3- [Hiv] didemnin A,

Z-didemnin A,

9- [Z-Nva] aplidine,

3- [Hiv] -9- [Z-ala] aplidine,

8- [Gly] -9- [coumarin] didemnin A,

8- [biotin] didemnin A,

3- [Hiv] -7.8- [spiro] -9- [Boc] aplidine,

3- [Hiv] -Z-didemnin A,

3- [Hiv] -9- [Z-Nva] aplidine,

7.8- [spiro] -9- [pyr] aplidine,

3- [Hiv] -9- [lac (OTBDMS)] aplidine,

3- [Hiv] -9- [Boc-Ala] aplidine,

7.8- [spiro] -9- [Boc] aplidine,

3- [Hiv] -8- [Boc-Val] aplidine,

8- [Val] -9- [isobutyryl] didemnin A,

3- [Hiv] -8- [hexanoyl] didemnin A,

3- [Val] -9- [Lac (OTBDMS)] aplidine,

3- [Aip] didemnin A,

3- [Hiv] -9- [D-Lac (OTBDMS)] aplidine,

7.8- [spiro] -9- [isobutyryl] aplidine,

3- [Hiv] -7.8- [spiro] -9- (Pyr] aplidine,

3- [Hiv] -7.8- [spiro] -9- [isobutyryl] aplidine,

3- [Hiv] -7.8- [spiro] -9- [acryloyl] aplidine or

[Aip] ^3- aplidine.

The present invention also relates to compounds having the following formulas:

and related structures.

In one particularly preferred embodiment, the present invention relates to a synthesis method with the formation of aa3 = [Hiv] ³ or [Val] ³ or [Aip] ³ , which are an integral part of a number of extremely effective and rare antitumor agents developed and claimed for clinical trials, after they have become available in adequate amounts, and their simplest isomers, in which the amino acid residues are rearranged. The specified method is enantio- and stereo-adjustable and fast, with the advantages of standard methods of synthesis methods in the solution phase.

A preferred embodiment of the present invention is described by Formula I, wherein aa3 independently represent α-amino acids of L or D configuration. If used, X independently means —C (R) ₂ -, O, S or NR; where R independently means H or an organic group selected from the group consisting of an alkyl group, an aryl group, an aralkyl group and derivatives thereof substituted with a hydroxy group, a mercapto group, an amino group, a guanidino group, a halogen group. Wherein R most preferably contains from about 1 to 12 carbon atoms, more preferably from about 1 to 8 carbon atoms and, even more preferably, from about 1 to 6 carbon atoms, most preferably 1, 2, 3 or 4 carbon atoms. Methyl, ethyl and propyl, including isopropyl, are particularly preferred alkyl groups in the compounds of the present invention. Where aa3 most preferably means α- (α'-hydroxyisovaleryl) propionyl (Hip), (X = O, Y = -COCHCH ₃ CO-) - series A, or α- (α'-aminoisovaleryl) propionyl (Aip) (X = NH, Y = -ССОСН ₃ СО-) - series N or valine (X = NH, Y = -СО-) - series V, or α-hydroxyisovaleryl (X = -O-, Y = -СО-) - series H, or N-methylvaline (X = NMe, Y = —CO—) —M series. Where aa8 independently mean α-amino acids of the L or D configuration, if used; where X ₂ independently means CR, O, S or N,

where R independently means H or an organic group selected from the group consisting of alkenyl, alkyl group, aryl group, aralkyl group and derivatives thereof substituted with hydroxy group, mercapto group, amino group, guanidino group, halogen group, where each of R ₁ , R ₂ , R ₃ and R ₄ independently mean H or an organic group selected from the group consisting of an alkyl group, an aryl group, an aralkyl group and derivatives thereof substituted with a hydroxy group, a mercapto group, an amino group, a guanidino group, a halogen group. Aa8 may also be a proline residue, as in formula II. Wherein R ₃ independently means H or an organic group selected from the group consisting of an alkyl group, an aryl group, an aralkyl group and derivatives thereof substituted with a hydroxy group, mercapto group, amino group, guanidino group, halogen group.

Wherein R _{3 can} most preferably be pyruvic acid, an aralkyloxycarbonyl group or amino acid, or peptides. Alkyl groups preferably have from about 1 to 12 carbon atoms, more preferably from about 1 to 8 carbon atoms, even more preferably from about 1 to 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms. Methyl, ethyl and propyl, including isopropyl, are particularly preferred alkyl groups in the compounds of the present invention. As used here, the term alkyl, unless otherwise indicated, refers to both cyclic and non-cyclic groups, even if cyclic groups consist of at least three carbon members of the cycle. Preferred amino acids are protected or unprotected D or L glycine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, methionine, cysteine, aspartate, asparagine, glutamic acid, glutamine, lysine, arginine, proline, serine, threonine, histroline and. Preferred peptides may be composed of the above amino acids.

In addition, aa8 and R ₄ can be linked via derivatives of 6-oxo-1,7-diazaspiro [4,4] nonane structures:

where R ₅ most preferably may mean pyruvic acid, an aralkyloxycarbonyl group or amino acid, or peptides. Alkyl groups preferably have from about 1 to 12 carbon atoms, more preferably from about 1 to 8 carbon atoms, even more preferably from about 1 to 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms. Methyl, ethyl and propyl, including isopropyl, are particularly preferred alkyl groups in the compounds of the present invention. As used here, the term alkyl, unless otherwise indicated, refers to both cyclic and non-cyclic groups, even if cyclic groups consist of at least three carbon members of the cycle. Preferred amino acids are protected or unprotected D or L-glycine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, methionine, cysteine, aspartate, asparagine, glutamic acid, glutamine, lysine, arginine, proline, serine, threonine, histroline and . Preferred peptides may be composed of the above amino acids.

As used here, the term "organic group" means a hydrocarbon group, divided into an aliphatic group, a cyclic group or a combination of aliphatic and cyclic groups (for example, aralkyl groups). In the context of the present invention, the term “aliphatic group” means a saturated or unsaturated linear or branched hydrocarbon. The term is used, for example, as encompassing alkyl, alkenyl and alkynyl groups. The term “alkyl group” means a saturated linear or branched hydrocarbon group including, for example, methyl, ethyl, isopropyl, isobutyl, tert-butyl, heptyl, dodecyl, octadecyl, amyl, 2-ethylhexyl, 2-methylbutyl, 5-methylhexyl and the like like that. The term “alkenyl group” means an unsaturated, linear or branched hydrocarbon group with one or more carbon-carbon double bonds, such as a vinyl group. The term "alkynyl group" means an unsaturated, linear or branched hydrocarbon group with one or more triple carbon-carbon bonds. The term “cyclic group” means a ring-closed hydrocarbon group, subdivided into an alicyclic group, an aromatic group or a heterocyclic group. The term "alicyclic group" means a cyclic hydrocarbon group having characteristics that make it similar to aliphatic groups. The term “aromatic group” or “aryl group” means a mono- or polycyclic aromatic hydrocarbon group. The term “heterocyclic group” means a ring-closed hydrocarbon group in which one or more atoms of the cycle mean an element other than carbon (for example, nitrogen, oxygen, sulfur, etc.).

Preferred alkoxy groups in the compounds of the present invention include groups having one or more oxygen bridges and from about 1 to 12 carbon atoms, more preferably from about 1 to 8 carbon atoms, even more preferably from about 1 to 6 carbon atoms, and most preferably 1, 2, 3 or 4 carbon atoms.

Suitable heteroaromatic groups in the compounds of the present invention contain one, two or three heteroatoms selected from N, O or S atoms, and include, for example, coumarin, including 8-coumarin, quinolinyl, including 8-quinolinyl, pyridyl, pyrazinyl, pyrimidyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl, indolyl, benzofuranyl and benzothiazole. Suitable heteroalicyclic groups in the compounds of the present invention contain one, two or three heteroatoms selected from N, O or S atoms, and include, for example, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholino and pyrrolidinyl groups.

Suitable carbocyclic aryl groups in the compounds of the present invention include mononuclear or polynuclear compounds, including polynuclear compounds that contain single and / or fused aryl groups. Typical carbocyclic aryl groups contain 1-3 individual or fused rings and from about 1 to 18 carbon atoms in the ring. Particularly preferred carbocyclic aryl groups include phenyl, including substituted phenyl, such as 2-substituted phenyl, 3-substituted phenyl, 2,3-substituted phenyl, 2,5-substituted phenyl, 2,3,5-substituted and 2,4 , 5-substituted phenyl, where one or more phenyl substituents represent an electron-attracting group such as halogen, cyano, nitro, alacanoyl, sulfinyl, sulfonyl and the like; naphthyl, including 1-naphthyl and 2-naphthyl; diphenyl; phenanthryl and anthracyl.

Optionally protected amino groups may be protected using groups known for the indicated purpose. Suitable protecting groups for amines include carbamates, amides, and other protecting groups such as alkyl, arylalkyl; sulfo- or halo-arylalkyl-; haloalkyl, alkilsililalkil-, arylalkyl, tsikloalkilalkil-, alkilarilalkil-, heterocycloalkyl-, nitroarilalkil-, atsilaminoalkil-, nitroarilditioarilalkil-, ditsikloalkilkarboksamidoalkil-, cycloalkyl, alkenyl, arylalkenyl-, nitroarilalkenil-, geterotsikloalkenil-, heterocyclo, gidroksigeterotsiklo- alkyldithio; alkoxy- or halogen- or alkylsulfinylarylalkyl; heterocycloacyl- and other carbamates, and alacanoyl-, haloalacanoyl-, arylalacanoyl-, alkenoyl-, heterocycloacyl-, aroyl-, arylaroyl-, halogenaroyl-, nitroaroyl-, alkyloalkylalkylalkylalkylalkylalkylalkyl , alkoxyarylalkyl, cycloalkyl, nitroaryl, arylalkyl, alkoxy or hydroxyarylalkyl and many other groups. Such groups may be optionally substituted with the above substituent groups.

When referring here to substituted groups in the compounds of the present invention, a specific radical is meant which may be substituted at one or more existing positions by one or more suitable groups, for example, halogen such as fluorine, chlorine, bromine and iodine; cyano; hydroxyl; nitro; azido; alacanoyl, such as a C1-6 alkanoyl group, such as acyl and the like; carboxamido; alkyl groups, including alkyl groups with about 1-12 carbon atoms or 1-6 carbon atoms, and more preferably 1-3 carbon atoms; alkenyl or alkynyl groups, including groups having one or more unsaturated bonds and about 2-12 carbon or 2-6 carbon atoms; alkoxy groups, including these groups having one or more oxygen bridges and about 1-12 carbon atoms or 1-6 carbon atoms; aryloxy such as phenoxy; alkylthio groups, including these groups having one or more simple thioether bonds and about 1-12 carbon atoms or 1-6 carbon atoms; alkylsulfinyl groups, including these radicals having one or more sulfinyl bonds and about 1-12 carbon atoms or 1-6 carbon atoms; alkylsulfinyl groups, including these radicals having one or more sulfonyl bonds and about 1-12 carbon atoms or 1-6 carbon atoms; aminoalkyl groups, such as groups having one or more N atoms and about 1-12 carbon atoms or 1-6 carbon atoms; carbocycloaryl with 6 or more atoms, in particular phenyl (for example, R is a substituted or unsubstituted diphenyl radical); and aralkyl, such as benzyl.

As is well known in the art, a high degree of substitution is not only permissible, but often appropriate. The compounds of the present invention may be substituted. In order to simplify the terminology used in this description, the terms “group” and “radical” are used to distinguish between chemical groups intended for substitution or which may be substituted, and corresponding groups for which substitution is not provided or which may not be substituted in this way. Thus, when the term “group” is used to describe a chemical substituent, an unsubstituted group is meant and this group with O, N or S atoms, for example, in a chain, as well as a carbonyl group or other standard substitution. When the term “radical” is used to describe a chemical compound or substituent, it is intended to include only an unsubstituted chemical structure. For example, the term “alkyl group” is intended to include not only fully open-chain saturated hydrocarbon alkyl substituents, such as methyl, ethyl, propyl, isobutyl and the like, but also alkyl substituents carrying additional substituents known in the art, such as hydroxy, alkoxy, amino, carboxyl, carboxamido, halogen atoms, cyano, nitro, alkylsulfonyl and so on. Thus, an “alkyl group” includes ether groups, haloalkyls, alcohols, thiols, carboxyl, amines, hydroxyalkyls, sulfoalkyls and so on. On the other hand, the expression “alkyl radical” is limited to the inclusion of fully saturated, fully open chain saturated hydrocarbon alkyl substituents such as methyl, ethyl, propyl, isobutyl and the like.

Other aspects of the present invention relate to synthetic methods.

The method involves obtaining a fragment of didemnin having the structure

the method comprises combining Boc-D-allo-Ileu-OH with benzyl acetate lithium enolate.

The carbonyl group of a didemnin fragment of the formula:

can be restored to obtain a fragment of didemnin of the formula:

The hydroxyl group of a compound of the formula:

can be protected to obtain a fragment of didemnin having the structure

Subsequent deprotection of the benzyl ester group gives a didemnin fragment having the structure

Another method of the present invention for producing a didemnin fragment comprises a combination of a first reagent having a structure

and a second reagent having the structure

leading to the formation of a fragment of didemnin having the structure

where X is selected from the group consisting of —O— and —NH—; where R is an amino protecting group and where R is a hydroxy protecting group. Conveniently, when X is —O— and R is tert-butyldimethylsilyl or X is —NH— and R is Boc.

Another method of the present invention for producing a didemnin fragment comprises a combination of a first reagent having a structure

and a second reagent having the structure

leading to the formation of a fragment of didemnin having the structure

where X is selected from the group consisting of —O—, NMe and —NH—; where R is an amino protecting group and where R is H. Conveniently when X is —O— and R is H; or X is —NH— and R is Boc; or X is —NMe— and R is Boc.

The method of the present invention includes hydrolysis of a fragment of didemnin of the formula:

leading to the formation of a fragment of didemnin having the structure

where X is selected from the group consisting of —OH and —NH ₂ .

Another method involves hydrolysis of a didemnin fragment

leading to the formation of a fragment of didemnin having the structure

where X is selected from the group consisting of —NH ₂ and —NHMe.

Another method for producing a didemnin fragment has been developed, the method comprising combining a first reagent having the structure

and a second reagent having the structure

leading to the formation of a fragment of didemnin having the structure

where X is selected from the group consisting of —O—, —NMe and —NH—; where Y means - (SOSCHN ₃ ) _n CO-; where n is 0 or 1.

The method, characterized in that it includes hydrolysis of a fragment of didemnin

leading to the formation of a fragment of didemnin having the structure

where X is selected from the group consisting of —O— and —NH—; where Y means - (SOSCHN ₃ ) _n CO-; where n is 0 or 1.

The present invention provides yet another method for producing a didemnin fragment, the method comprising combining a first reagent having a structure

and a second reagent having the structure

leading to the formation of a fragment of didemnin having the structure

The method of the present invention includes removing the protective group from the benzyl ester group of a didemnin fragment of the formula

leading to the formation of a fragment of didemnin having the structure

According to the present invention, a method for producing a didemnin fragment comprises a combination of a first reagent having a structure

and a second reagent having the structure

leading to the formation of a fragment of didemnin having the structure

where X is selected from the group consisting of —O— and —NH—; where Y means - (SOSCHN ₃ ) _n CO-; where n is 0 or 1.

The method includes deprotecting a fragment of didemnin

leading to the formation of a fragment of didemnin having the structure

where X is selected from the group consisting of —O— and —NH—; where Y means - (SOSCHN ₃ ) _n CO-; where n is 0 or 1.

Another method of the present invention for producing a didemnin fragment involves cyclizing a fragment of the formula:

leading to the formation of a didemnin analog having the structure

where X is selected from the group consisting of —O— and —NH—; where Y means - (SOSCHN ₃ ) _n CO-; where n is 0 or 1.

The method includes hydrolysis of a didemnin analog

leading to the formation of a didemnin analog having the structure

where X is selected from the group consisting of —O— and —NH—; where Y means - (SOSCHN ₃ ) _n CO-; where n is 0 or 1.

Additionally, a method for producing a didemnin analog has been developed, the method includes combining a first reagent having a structure

and a second reagent having the structure

leading to the formation of a didemnin analog having the structure

where X is selected from the group consisting of —O— and —NH—; where Y means - (SOSCHN ₃ ) _n CO-; where n is 0 or 1.

Another method involves deprotecting a fragment of a didemnin formula:

leading to the formation of a fragment of didemnin having the structure

where X is selected from the group consisting of —O— and —NH—; where Y means - (SOSCHN ₃ ) _n CO-; where n is 0 or 1.

A method for producing a didemnin fragment comprises a combination of a didemnin fragment having the structure

and a second reagent having the structure

leading to the formation of a fragment of didemnin having the structure

Another method involves deprotecting a fragment of a didemnin formula:

leading to the formation of a fragment of didemnin having the structure

The method of the present invention for producing a didemnin analog comprises a combination of a didemnin analog of the formula:

with fragment

leading to the formation of a didemnin analog having the structure

where X is selected from the group consisting of —O— and —NH—; where Y is - (COCHCH ₃ ) _n CO—; where n is 0 or 1.

A method of obtaining a didemnin analog includes a combination of a didemnin analog having the structure:

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

where X is selected from the group consisting of —O— and —NH—, and R is isopropyl; where X is —O— and R is n-propyl and R is n-pentyl.

The method of the present invention for producing a didemnin analog includes a combination of a didemnin analog having the structure:

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

Where:

The method of the present invention includes deprotecting a didemnin analog

leading to the formation of a fragment of didemnin having the structure

Where:

A method has been developed for the preparation of a didemnin analog, comprising a combination of a didemnin analog having a structure

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

The method further includes removing the protection of the analog didemnin

leading to the formation of a didemnin analog having the structure

A method for producing a didemnin analog has been developed, including a combination of a didemnin analog

and isobutyryl chloride, leading to the formation of a didemnin analog having the structure

A method has been developed for the preparation of a didemnin analog, comprising a combination of a didemnin analog having a structure

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

where R is Boc, isobutyryl, pyruvyl or acryloyl.

A method of producing a didemnin analog includes a combination of a didemnin analog having a structure

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

where R is SO ₂ Me or Z-Nva.

A method of the present invention has been developed, including deprotection of a didemnin analog

leading to the formation of a didemnin analog having the structure

where R ₂ means Nva.

An integral part of the present invention is a method for producing a didemnin analog, comprising a combination of a didemnin analog having a structure

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

where R is Boc, isobutyryl or pyruvyl.

A method of producing a didemnin analog includes a combination of a didemnin analog having a structure

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

A method has been developed for the preparation of a didemnin analog, comprising a combination of a didemnin analog having a structure

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

A method of producing a didemnin analog comprising a combination of a didemnin analog having a structure

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

A method has been developed for the preparation of a didemnin analog, comprising a combination of a didemnin analog having a structure

and methylsulfonyl chloride, leading to the formation of a didemnin analog having the structure

A method of obtaining a didemnin analog includes a combination of a didemnin analog having a structure

and a fragment having a structure

leading to the formation of a didemnin analog having the structure

where X is O and S, where R is butyl and phenyl.

It should be understood that all of these methods are illustrative of the invention and, if desired, can be modified. In particular, other protecting groups may be used to protect amino groups or hydroxy groups. Other reagents may be used to administer the intended groups. If desired, substituents can be varied in a particular embodiment of the general formula of the compounds of this invention and examples of preferred substituent values. Modified methods are an integral part of the present invention.

In order to ensure that this description to the necessary extent includes all disclosures of priority patent applications and to ensure the right of declared priorities in accordance with the filing dates, applicants include in the description, by reference, the contents of patent applications GB 0016148.9 and GB 0103750.6.

Compounds of the present invention can be obtained by synthetic methods. The methods for synthesizing aplidine and derivatives described herein can also be used to synthesize a wide variety of didemnins.

The structures of some compounds are shown in figure 1:

Derivatives of aa3-aplidine are synthetic cyclic depsipeptides structurally similar to aplidine (compound II) (aa3 = Hip, also known as dehydrodidemnin B), which is a natural didemnin isolated from ascidium compounds Aplidium albicans (figure 1). The resulting molecules are distinguished by the presence of hydroxyisovaleric acid (Hiv) (compound I), or valine (compound V), or methylvaline (compound VI), or α- (α'-aminoisovaleryl) propionyl (Aip) (compound VII), instead of hydroxyisovalerylpropionic (Hip ) (II) a link which is present in all other compounds of a natural origin belonging to the class of didemnin. A similarity of these two structures has also recently been found when comparing didemnin B (compound III), the most well-known member of the indicated class of depsipeptides, and a new cyclic depsipeptide isolated from unidentified compounds of the Brazilian ascidia of the didemnid family: tamandarin A (compound IV). Compounds I, V, VI, and VII are unnatural derivatives of didemnin.

Structural homology between III and IV is also reflected in their corresponding biological activity. Comparison of both compounds showed that IV retains the same levels of antitumor activity in vitro in clonogenic trials, as well as inhibitory properties in relation to protein biosynthesis, and is slightly more active in vitro than III against pancreatic carcinoma. However, compound IV shows absolutely no specificity for any type of NCI 60-cell panel tumors. Didemnin B has been shown to be toxic at doses close to those required for therapeutic use, and IV seems to be a wide-spectrum toxin rather than a selective agent.

[Hiv] ^3- aplidine (I), on the contrary, has the same advantages as those found for aplidine (II) with respect to didemnin B (III), that is, it is more specific with respect to solid tumors such as a colon tumor, chondrosarcoma or osteosarcoma according to the MTS test. In contrast, [Val] ^3- aplidine (V) and [MeVal] ^3- aplidine (VI) are new compounds with a high level of antitumor activity in vitro. Finally, compounds V, VI, and VII cause, obviously, in comparison with the starting aplidine, increased formation of hydrogen bonds in the active region, and thus represent more active compounds. In addition, the existing replacement of the amide bond with an ester bond can increase the stability of the cyclodepsipeptide core.

Here, the basic general synthesis of various series of aplidine derivatives is described. As an example, figure 2 shows a retrosynthetic analysis.

Key steps include efficient macrocyclization of linear precursors 6, convenient stereoselective synthesis of the Ist-aa3-Leu-Pro link (A1) and the right fragment D1. The final compound of macrocycle 4 with different side chains gives aa3-aa8-aplidine and derivatives. The reliability of this synthetic approach has been successfully proven in the development of practical synthesis of aplidine II (aa3 = Hip).

The formation of a macrocyclic core is the main key stage in all series. In previous syntheses of didemnins, the successful cyclization of all four possible amide bonds was achieved. However, in this work, the formation of the N (Me) -O (Me) -Tyr and Pro bond was chosen as the moment of macrocyclization, which is based on previous studies related to the general synthesis of aplidine II (G. Jou, I. Gonzalez, F. Albericio, P. Lloyd-Williams, E. Giralt., J, Org. Chem. 1997, 62, 354-366 and Patent ES-2102322).

The macrocyclic core of the target molecule is divided into the left link of the tetrapeptide Ist-aa3-Leu-Pro A1 and the dipeptide Boc-Thr- (Z-NMe-OMe-Tyr) -OH D1.

The synthesis of the right fragment of dipeptide D1 has already been described. However, the synthesis, schematically depicted by scheme 1, allows to obtain this intermediate product in kilogram scales. The acid functionality of Boc-Thr (OH) -OH is protected by phenacyl bromide, which gives directly the alcohol D3, which is esterified with Z-N (Me) -O (Me) -Tyr-OH using DCC in the presence of DMAP to give D2. Removal of the phenacyl group with Zn in acetic acid gives a pure D1 fragment.

Obtaining the precursor A1 for the various series is shown schematically in Scheme 2. For the SHPL, SVPL, and SMPL series, the first stage of the combination of Leu-Pro-OBzl (A5) and industrially produced acid (B1) gives directly alcohol A3, ready for the next reaction with isostatin (C1 ) For the synthesis of SAPL and SNPL series, the method differs from the above in that B1 is a β-keto ester synthesized from [α- (α'-hydroxyisovaleryl) propionic and [α- (α'-aminoisovaleryl) propionic acid, respectively. The synthesis of fragment B2 (SAPL and SNPL series) is shown schematically in Scheme 3. Hydrogenolysis with the conversion of B2 to B1 is carried out immediately before (in a single reactor) coupling reaction with A5.

The preparation of the complete fragment A1 depends on the availability of the isostatin part, which is obtained from nonproteinogenic amino-protected Boc-D-alloisoleucine (C5) in large quantities. A synthetic method for producing C1 is shown schematically in scheme 4.

Activation of the carboxyl functionality of Boc-D-allo-Ile by carbonyldiimidazole followed by condensation of benzyl acetate with lithium enolate affords the β-keto ester (C4). Stereoselective reduction using KVN ₄ in methanol gives C3. Protection of the secondary hydroxyl group in the form of a simple TBDMS ether (C2) and hydrogenolysis of the resulting benzyl ester give isostatin (C1).

The subsequent steps for preparing compounds I, II, V, and VII are depicted in Scheme 5.

The binding of fragments A1 and D1 to HBTU / HOBt gives a linear precursor of type 7. The hydrogenolysis of Cbz and benzyl protecting groups proceeds cleanly and calmly with Pd (OH) ₂ , which gives 6. The macrocyclization step using HATU / HOAt gives intermediate 5 with a good yield (75%). The hydrogen chloride in dioxane is used to cleave the Boc-protecting group leading to amine 4. This compound is coupled to Z-NMe-D-Leu-OH to give 3, which is subjected to hydrogenolysis under the influence of Pd-C. The resulting compound 2 was coupled to the Pyr-Pro-OH side chain using DIPCDI to give the corresponding compounds.

Interestingly, type 2 precursors, which are analogues in all of the above relations of didemnin A, serve as the starting blocks used in the synthesis to construct some of the compounds of this class in question, since the N-terminal, free secondary amino group is the site of attachment of various acyl groups to the cyclic depsipeptide.

Earlier studies (Rinehart et al., J. Med. Chem, 1996, 39, 2819-2834) also found that the acyl derivatives of didemnin A (dA) 3 are active as starting compounds 2. For the aa3 series, applicants also found activity in the compounds 3.

Tables 1 and 2 show the IC ₅₀ values found for compounds 1.

Table 1
Cytotoxicity of compounds of the type [AA3] -aplidine IC ₅₀ (molar) Compound / Series P388 A549 NT29 MEL28 Aplidine (II) / SAPL1 1.80E-10 1.80E-10 4,50Е-10 4,50Е-10 [Hiv] ^3- aplidine (I) / SHPL1 4.74E-10 4.74E-10 4.74E-10 4.74E-10 [Val] ^3- aplidine (V) / SVPL1 1,13E-10 1,13E-10 1,13E-10 1,13E-10 [Aip] ^3- aplidine (VII) / SNPL1 9.01E-10 9.01E-10

Methodology: according to Berjeron et al., Biochem and Bioph Res, Comm., 1984, 121, 3, 848-854.

P388 = mouse lymphoma. A549 = human lung carcinoma. HT29 = human colon carcinoma. MEL28 - human melanoma.

Table 2 IC ₅₀ Values (Molar) for the Aplidine Family Didemnin
B9LSAPL1 Aplidine SAPL1 [Hiv] ^3- Aplidine SHPL1 Solid tumors Line III II I Bladder 5637 2.50E-08 3.59E-08 9.02E-08 Breast MX-1 1.54E-06 1.67E-07 H / a Colon or colon HT-29 8.07E-08 6.87E-07 1.02E-08 Stomach Hs746t 6.60E-09 2.52E-08 7.16E-08 Liver SK-HEP- 9.21E-08 9.44E-08 2.66E-07 one NSCLC A549 1.21E-04 2.40E-05 H / a Ovary SK-OV-3 1.63E-07 7.20E-08 - Pancreas PANC-1 1.52E-10 1.7E-07 - Pharynx Fadu 9.79E-08 7.29E-08 3.71E-08 Prostate PC-3 9.00E-08 5.13-07 - Prostate DU-145 - - H / a Prostate Lncap - - 1.46E-08 Kidney 786-o 2.90E-07 8.31 E-08 - MRL NCl- - - H / a H187 Retinoblastoma Y-79 - - Melanoma Mel-28 - - 4.86E-07 Fibrosarcoma SW 694 3.28E-06 1.49E-06 H / a Chondrosarcoma CHSA - 3.45E-09 Osteosarcoma OSA-FH - - 5.89E-09

Didemnin B Aplidine [Hiv] ³ - Leukemia / Lymphoma Line 9LSAPL1 SAPL1 Aplidine III SHPL1 II I ALL (promyelocytic leukemia) Hl-60 1.44E-07 7.89E-08 ON ALL (acute lymphoblastic) Mott 3 5.45E-07 5.95E-07 1.77E-08 CML (chronic myelogenous) K562 3.31E-06 5.72E-07 5.21E-07 ALL (B-cell) CCRF-SB 6.55E-07 4.72E-07 - Leukemia (capillary-B-keto) Mo-b - - - Leukemia (plasma cell) ARH-77 - 1.78E-07 - Lymphoma (T-cell) H9 2.13E-07 5.25E-07 ON Lymphoma (skin T cells) Hut 78 3.56E-08 4.47E-08 - Lymphoma (undifferentiated) MC116 8.84E-09 9.21E-07 3.82E-07 Lymphoma (Burkitt B cells) RAMOS - - - Lymphoma (histiocytic) U-937 1.87E-07 5.62E-07 - Lymphoma (B-cell) Mob - - - Lymphoma (Burkitt ascites) P3HR1 5.58E-08 5.34E-08 - Methodology: MTT MTT MTS (new) N / A = not active

The compounds obtained here are shown in Schemes 5 and 6. These compounds include N ^α -propionyl- [aa] ³ -dA, N ^α -butyryl- [aa] ³ -dA, N ^α -isobutyryl- [aa] ³ -dA and N ^α- pentanoyl- [aa] ³ -dA.

Analogues having two acyl substituents after the N-ends of the aa3-dA nucleus were obtained to study the influence of structural factors on specificity for some tumors. Diacyl compounds, isobutyryl-Pro-OH, O-isobutyryl-Lac-Pro-OH, N-benzyl-Ala-Pro-OH, were obtained and condensed with type 2 compounds by the DIPCDI method, which yields, respectively, after deprotection and purification, [aa] ³ - [isobutyryl] ^9- aplidine, [O-isobutyryl-Lac] ^9- aplidine, [Ala] ^9- aplidine.

Preferred aplidine derivatives: compounds of the series: H, V

The compounds also bind to give active compounds:

The synthesis of spirocyclic fragments is shown schematically in Scheme 7. The synthesis is based on the previously described compound 8. References: a) Seebach, D. et al. J. Am. Chem. Soc. 1983, 105, 5390-5398. b) Genin, M. J. et al. J. Org. Chem. 1993, 58, 2334-2337.

Preferred aplidine derivatives: Series A compounds

Spiro compounds are also linked, as in the previous series, to obtain active compounds:

Spiro = [(5R) -1- (substituent at position 9) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane] ^7-9

(1) This compound is described in the literature: Joc Org. Chem. 2000, 65, 782-792. Their synthesis was published before their discovery (j): Org. Lett., 2000, vol 0, No. O, A-D.

Methodology: according to Berjeron et al, Biochem and Bioph Res. Comm., 1984, 121, 3, 848-854

388 = mouse lymphoma. A549 = human lung carcinoma. HT-29 = human colon carcinoma. MEL-28 - human melanoma. DU145 - carcinoma of the human prostate gland.

388 = Murine lyrnphoma. A549 - human lung carcinoma. HT-29 - human colon carcinoma. MEL-28 - human melanoma. DU145 = human prostate carcinoma

List of Symbols Miscellaneous AA Amino acid Isst Isostatin Hip Hydroxyisovalerylpropionic acid Hiv Hydroxyisovaleric acid Aip Aminoisovalerylpropionic acid Lac Lactic acid LC Liquid Chromatography (LC) HPLC High Performance Liquid Chromatography (HPLC) TLC Thin layer chromatography (TLC) M.p. Melting point (mp.) R _t Retention time Quant. Quantitative yield (count) ESI-MS Mass spectrum of ionization electrospray Protective groups Bn Benzyl Boc tert-butyloxycarbonyl TBDMS tert-butyldimethylsilyl Z Benzyloxycarbonyl Ras Phenylacetyl Solvents Thf Tetrahydrofuran (THF) Hex Hexane ACN Acetonitrile DCM Dichloromethane EtOAc Ethyl acetate DMF Dimethylformamide (DMF) MTBE Methyl tert-butyl ether Et ₂ O Diethyl ether t-buoh tert-butanol TFA Trifluoroacetic acid Meoh Methanol EtOH Ethanol IPA Isopropanol Reagents Cdi 1,1'-Carbonyldiimidazole Hobt 1-hydroxybenzotriazole HBTU N-oxide N - [(1H-Benzotriazol-1-yl) (dimethylamino) methylene] -N-methanammonium hexafluorophosphate BOP-C1 Bis (2-oxo-3-oxazolidinyl) chloride phosphinic acid Hatu N-oxide N - [(dimethylamino) -1H-1,2,3, -triazolo [4,5-b] pyridin-1-ylmethylene] -N- methyl methanammonium hexafluorophosphate HOAt 1-Hydroxy-7-aza-benzotriazole DCC Dicyclohexylcarbodiimide Dipcdi N, N'-Diisopropylcarbodiimide Tbaf Tetrabutylammonium fluoride AcOH Acetic acid p-TsOH P-toluenesulfonic acid DMAP 4-dimethylaminopyridine NMM N-methylmorpholine Dipa Diisopropylethylamine Tea Triethylamine TFA Trifluoroacetic acid

General methodology

All work is performed in an inert atmosphere of argon. All solvents have reagent purity (used for processing) or HPLC purity (used as solvent for reaction or purification). Anhydrous solvents are used directly as they go on sale. Tetrahydrofuran is freshly distilled before use to remove the stabilizer. All other reagents are commercially available compounds of the highest available purity. Analytical thin layer chromatography (TLC) was carried out on aluminum plates with Merck silica gel (60, F254), previously coated with a fluorescent indicator. Visualization is carried out using ultraviolet light (254 nm), phosphomolybdenum acid (5% m / v) in 95% ethanol or vanillin. The proton or carbon magnetic resonance spectrum ( ¹ H, ¹³ C-NMR) is recorded on a Varian-300 (300 MHz) Fourier transform spectrometer, and chemical shifts are expressed in parts per million (ppm) relative to CHCl ₃ as an internal standard (7.26 ppm for ¹ H and 77.0 for ¹³ C). Multiplets are designated as singlet (s), doublet (d), doublet of doublets (dd), doublet of triplets (dt), triplet (t), quartet (q), multiplet (m) and widened singlet (us.s), and constants interactions (J) are expressed in Hz. Optical rotations (in degrees) are measured using a Jasco P1020 polarimeter. Electron Spray Ionization Mass Spectrum (ESI-MC) is acquired on the Hewlett Packard Series 1100 MSD. Elemental. Flash column chromatography was performed on E. Merck silica gel 60 (240-400 mesh) or RP C18 (40-63 μm) using the solvent systems indicated in separate experiments.

The following procedures describe the synthesis of intermediates derived from aplidine (SAPL), [Aiv] ^3- aplidine (SNPL), [Hiv] ^3- aplidine (SHPL), [Val] ^3- aplidine (SVPL) and [MeVal] ³ - aplidine (SMPL).

Example 1

Synthesis of Benzyl (4R, 5S) -4- (tert-butoxycarbonylamino) -5-methyl-3-oxoheptanoate (C4)

To a solution of Boc-D-alloIle-OH (15.26 g, 65.9 mmol) in dry THF (200 ml) at 0 ° C. under argon atmosphere was added CDI (16.04 g, 98.96 mmol). After 15 minutes, the mixture was allowed to warm to room temperature and stirred for 16 hours. The resulting solution was cooled to -78 ° C and added via cannula to a well-stirred solution of benzyl lithium enolate cooled to -78 ° C (625 ml, 0.37 M), [obtained by dropwise adding a solution of benzyl acetate (33.34 ml) in THF (165 ml) to a solution of lithium diisopropylamide (0.36 M) in THF / hex. 3: 1 (642 ml) at -78 ° C]. The temperature should be maintained <-75 ° C. The reaction mixture was stirred at -78 ° C for 60 minutes. Then the mixture was allowed to warm to -10 ° C (30 min), cooled again to -78 ° C and quenched with saturated aq. ammonium chloride (200 ml), and then extracted with DCM (3 × 500 ml) at room temperature. The combined extracts were washed successively with aq. us. NaHCO ₃ (500 ml) and brine (200 ml). Drying (Na ₂ SO ₄ ) followed by removal of the solvent gave an oil that was applied to C18 silica and filled to the top of the LC-RPC18 [Lichroprep RFC-18 (40-60 microns) column. Elution using a gradient of ACN-H ₂ O (60-100% ACN)] gives the product C4 as a colorless oil (16.7 g, 70%). [α] ²⁰ _D - 20.0 (s 1, CHCl ₃ ), TLC: Rf = 0.32 (Merck, RP-C18, ACN-H ₂ O 7: 3).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.77 (d, 3H), 0.94 (t, 3H), 1.25 (s, 9H), 1.60 (m, 1H), 1, 90 (m, 2H), 3.58 (s, 2H), 4.47 (dd, 1H), 5.00 (d, 1H), 5.18 (s, 2H), 7.35 (br s , 5H).

Example 2

Synthesis of (3S, 4R, 5S) -N- (tert-butoxycarbonyl) isostatin benzyl ester (C3)

C4 (16.7 g, 45.9 mmol) was dissolved in methanol (690 ml) at 0 ° C. Potassium borohydride (7.43 g, 137.8 mmol) was added to the stirred solution and after 30 minutes the reaction was quenched with aq. HCl (0.1 N), to pH 4, and the mixture was extracted with DCM (300 ml). The extract was washed successively with aq. NaHCO ₃ (100 ml, sat.) And brine (100 ml). Drying (Na ₂ SO ₄ ) followed by removal of the solvent gave C3 alcohol (15.7 g, 93%) as a colorless oil. Rf = 0.45 (hex. - EtOAc 2: 1); [α] _D = -9.5 (c 0.76, CHCl ₃ ); Rf = 0.45 (EtOAc-hex. 1: 2).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.81 (d, 3H), 0.90 (t, 3H), 1.20 (m, 1H), 1.36 (m, 1H), 1, 40 (s, 9H), 1.90 (m, 1H), 2.55 (dd, 1H), 2.70 (dd, 1H), 3.20 (d, 1H), 3.61 (m, 1H ), 3.90 (m, 1H), 4.40 (d, 1H), 5.18 (s, 2H), 7.40 (br s, 5H).

Example 3

Synthesis of Boc- (3S, 4R, 5S) -Ist (TBDMS) -OBn (C2)

To a solution of C3 (15.7 g, 42.9 mmol) in dry DMF (65 ml) at 0 ° C, imidazole (8.77 g, 128.8 mmol), DMAP (1.57 g, 12.81 mmol) are added. ) and TBDMS-C1 (19.42 g, 128.8 mmol). The reaction mixture was allowed to warm to room temperature overnight, then partitioned between Et ₂ O (200 ml) and successively aq. HCl (100 ml, 0.1 N) aq. NaHCO ₃ (100 ml, sat.) And brine (50 ml). After drying (Na ₂ SO ₄ ) and removal of solvent, the residue was purified by flash LC (silica gel, hex.) To give C2 (19.96 g, 97%).

[α] _D = 10.6 (c 1.01, CHCl ₃ ); Rf = 0.73 (EtOAc-Hex. 1: 2).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.15 (s, 6H), 0.82 (s, 9H), 0.85 (d, 3H), 0.89 (t, 3H), 1, 18 (m, 1H), 1.35 (m, 1H), 1.41 (s, 9H), 1.77 (m, 1H), 2.45 (dd, 1H), 2.60 (dd, 1H ), 3.62 (m, 1H), 4.20 (m, 1H), 4.40 (d, 1H), 5.05 (d, 1H), 5.15 (d, 1H), 7.40 (br.s, 5H).

Example 4

Synthesis of Boc- (3S, 4R, 5S) -Ist (TBDMS) -OH (Cl)

(с 0,594, DCM) [Лит. [α]_D 1,74 (с 2,64, CHCl₃). Synthesis, 1991, 294]; Rf=0,45 (Merck HPTLC, RP-C18, ACN-H₂O 8:2).To a solution of C2 (10.48 g, 21.8 mmol) in THF (110 ml) degassed and purged with argon was added Pd / C 10% (2.096 g, 20 wt.%). The mixture was stirred in an H ₂ atmosphere (1 atm) for 16 hours, then filtered through a 0.45 mm Teflon filter and concentrated under reduced pressure to obtain 7.8 g of a colorless oil. Colorless crystals (6 g, 70%) were obtained after crystallization in ACN at -20 ° C.

(from 0.594, DCM) [Ref. [α] _D 1.74 (c 2.64, CHCl ₃ ). Synthesis, 1991, 294]; Rf = 0.45 (Merck HPTLC, RP-C18, ACN-H ₂ O 8: 2).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.18 (s, 6H), 0.82 (s, 9H), 0.85 (d, 3H), 0.89 (t, 3H), 1, 10-1.20 (m, 2H), 1.42 (s, 9H), 1.80 (m, 1H), 2.50 (m, 2H), 3.58 (m, 1H), 4.11 (m, 1H).

Example 5

Synthesis of (2S) -2-tert-Butyl (dimethyl) silyloxy-3-methylbutyric acid (SAPLB4)

Imidazole (27.24 g, 400.11) is added to a stirred solution of (S) -2-hydroxy-3-methylbutyric acid (21.12 g, 181.9 mmol) in DMF (91 ml) at 0 ° C under argon atmosphere. mmol) and DMAP (6.94 g, 54.56 mmol). After 5 minutes, tert-butyldimethylchlorosilane (60.31 g, 400.11 mmol) was added. The mixture was allowed to warm to 23 ° C and stirred overnight. The crude reaction mixture was partitioned between Et ₂ O (250 ml) and aq. HCl (250 ml, 0.1 N). The organic phase is washed successively with aq. NaHCO ₃ (250 ml, sat.) And brine (250 ml), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to obtain a bis-like product as a pale yellow oil. A solution of the indicated product in THF (100 ml) was added dropwise (10 min) to a cooled (0 ° C) solution of KOH (30.47 g, 543 mmol) in THF / H ₂ O (543 ml, 181 ml). After 40 minutes, the reaction mixture was partitioned between H ₂ O (300 ml) and Et ₂ O (500 ml). The aqueous phase is partitioned between chilled (0 ° C.) aq. HCl (200 ml, 3n) and EtOAc (5 × 250 ml). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give SAPLB4 as a pale yellow oil (38.38 g, 91%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.02 (m, 6H), 0.90 (m, 15H), 2.08 (m, 1H), 4.06 (d, 1H).

Example 6

Synthesis of Benzyl- (4S) -4-tert-butyl (dimethyl) silyloxy-5-methyl-3-oxohexanoate (SAPLB3)

To a solution of (S) -2- (tert-butyldimethylsilyloxy) -3-methylbutyric acid (15.31 g, 65.9 mmol) in dry THF (200 ml) at 0 ° C. under argon, CDI (16.04 g 98.96 mmol). After 15 minutes, the mixture was allowed to warm to room temperature and stirred for 16 hours. The resulting solution was cooled to -78 ° C and benzyl lithium enolate (625 ml, 0.37 M) was added through a cannula to a well-stirred solution cooled to -78 ° C. , [obtained by adding dropwise a solution of benzyl acetate (33.34 ml) in THF (165 ml) to a solution of lithium diisopropylamide (0.36 M) in THF / hex. 3: 1 (642 ml) at -78 ° C]. The temperature should be maintained <-75 ° C. The reaction mixture was stirred at -78 ° C for 60 minutes. Then the mixture was allowed to warm to -10 ° C (30 min), cooled again to -78 ° C and quenched with aq. ammonium chloride (200 ml, sat.), and then extracted with DCM (3 × 500 ml) at room temperature. The combined extracts were washed successively with aq. NaHCO ₃ (500 ml, sat.) And brine (200 ml). Drying (Na ₂ SO ₄ ) followed by removal of the solvent gave an oil that was layered on C18 silica and filled on top of the LC-RPC18 [Lichroprep RPC-18 (40-60 microns) column. Elution using a gradient of ACN-H ₂ O (60-100% ACN)] gives SAPLB3 as a colorless oil (16.1 g, 70%). [α] _D - 25.0 (s 0.5, MeOH); Rf = 0.32 (Merck, RP-C18, ACN-H ₂ O 7: 3).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.02 (m, 6H), 0.92 (m, 15H), 1.92 (m, 1H), 3.63 (s, 2H), 3, 80 (s, 2H), 3.38 (d, 1H), 5.17 (d, 1H), 5.20 (d, 1H), 7.35 (br s, 5H).

Example 7

Synthesis of Benzyl- (4S) -4-tert-butoxycarbonylamino-5-methyl-3-oxohexanoate (SNPLB3)

Following the procedure developed for the synthesis of SAPLB3, from Boc-Val-OH (10 g, 46.0 mmol), the title compound is obtained after purification by flash LC (silica gel, hex gradient. EtOAc 10: 1 - 5: 1) as an oil (6.9 g, 43%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.77 (d, J = 7, 3H), 0.98 (d, J = 7 3H), 1.44 (s, 9H), 2.22 ( m, 1H), 3.58 (s, 2H), 4.31 (m, 1H), 5.03 (m, 1H), 5.18 (s, 2H), 7.34 (br s, 5H )

ESI-MC Calculate. for C ₁₉ H ₂₇ NO ₅ : 349.19. Found (m / z): 372.1 (M + Na) ⁺ .

Example 8

Synthesis of Benzyl- (2RS, 4S) -4-tert-butyl (dimethyl) silyloxy-2,5-dimethyl-3-oxohexanoate (SAPLB2)

SAPLB3 ester (15.12 g, 41.49 mmol) in dry THF (43 ml) was added dropwise to a solution of lithium diisopropylamine at -78 ° C [obtained by adding dropwise butyl lithium (1.6 M solution in hex .; 31, 12 ml, 49.79 mmol) to diisopropylamine (7.26 ml, 51.86 mmol) in dry THF (83 ml) in an Ar atmosphere at -78 ° C for 30 min]. The mixture was stirred for 0.5 h and then iodomethane (52.11 ml, 829.8 mmol) was added. The mixture was allowed to warm to 23 ° C and then stirring was continued for 24 hours. An additional amount of iodomethane (2.67 ml, 42 mmol) was added and the mixture was stirred for another 24 hours or until the starting material disappeared. Then the mixture was partitioned between aq. NH ₄ Cl (50 ml, sat.) And EtOAc (2 × 200 ml). The organic layer was washed successively with aq. NaHCO ₃ (100 ml, sat.), Brine (100 ml), dried (Na ₂ SO ₄ ), filtered and concentrated to give a yellow oil (12 g). The pure product (SAPLB2) is obtained after purification by LC on silica with a hex gradient. - Et ₂ O 100: 0 - 100: 2 in the form of a diastereomeric mixture of epimers on C2 (10 g, 63%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.01 (s, 3H), 0.02 (s, 3H), 0.91 (m, 15H), 1.30 (d, 3H), 2, 01 (m, H), 4.01 (m, 1H), 5.10 (d, 1H), 5.15 (d, 1H), 7.34 (br s, 5H).

Example 9

Synthesis of Benzyl- (2RS, 4S) -4-tert-butoxycarbonylamino-2,5-dimethyl-3-oxohexanoate (SNPLB2)

Following the procedure developed for the synthesis of SAPLB2, starting from SNPLB3 (10 g, 46.0 mmol), the title compound is obtained after purification by flash LC (silica gel, hex gradient - EtOAc 10: 1 - 5: 1), as diastereomeric mixtures (1: 1) of epimers on C2 (4.4 g, 62%). Rf = 0.4 and 0.37 (silica, hex. / EtOAc 3: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.72 (m, 3H), 0.86 (m, 3H), 1.37 (m, 3H), 1.44 (s, 9H), 2, 22 (m, 1H), 3.79 (m, 1H), 4.43 (m, 1H), 5.02 (m, 1H), 5.16 (m, 2H), 7.34 (m, 5H) )

ESI-MC Calculate. for C ₂₀ H ₂₉ NO ₅ : 363.29. Found (m / z): 364.1 (M + H) ⁺ .

Example 10

Synthesis of Leu-Pro-OBn as the hydrochloride salt (A5)

A solution of hydrogen chloride in dioxane (209 ml, 5.3 N) was added to a flask containing Boc-Leu-Pro-OBn (113.8 g, 272 mmol) and stirring was continued for 5 h or until complete conversion by TLC (disappearance starting material: Rf = 0.47 (hex. - EtOAc 2: 1, silicon dioxide). The solution was concentrated under reduced pressure and the resulting oil was distilled with CHCl ₃ (3 × 50 ml), CHCl _3- MTBE (30 ml - 50 ml ), MTBE (50 ml) and hex. (50 ml). The residue was dried in vacuo (16 h) to remove residual HCl to give the title compound as a white solid. A5 (96.4 g, 100%) was used without delay GOVERNMENTAL without further purification in the next stage, [α] _D ²² - 85.21 (c = 1, CHCl _3).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.92 (d, J = 7.1, 3H), 0.96 (d, J = 7.1, 3H), 1.55 (m, 1H) 1.82-2.14 (m, 5H), 2.26 (m, 1H), 3.42 (m, 1H), 3.90 (m, 1H), 4.32 (br s, 1H) ), 4.64 (m, 1H), 5.01 (d, J = 11.5, 1H), 5.16 (d, J-11.5, 1H), 7.34 (m, 5H), 8.40 (br s, 3H).

ESI-MC Calculate. for C ₁₈ H ₂₆ N ₂ O ₃ : 318.19. Found (m / z): 319.2 (M + H) ⁺ .

Example 11

Synthesis of TBDMS-Hip-Leu-Pro-OBn (SAPLA4)

Into a flask containing a degassed solution of SAPLB2 (20 g, 52.88 mmol) in THF bezv. (158 ml) equipped with gas inlet / outlet tubes, add 10% Pd / C (6.0 g, 30 wt.%) In an Ar atmosphere. Then, a stream of hydrogen is passed through the mixture for 8 hours or until complete conversion, according to TLC (disappearance of starting material). Ar was bubbled through the resulting mixture to displace hydrogen, and the mixture was filtered in an Ar atmosphere on a sintered glass funnel through a celite pad into a chilled flask (-5 ° С) containing HOBt (7.17 g, 52.88 mmol) and HBTU (21, 0 g, 55.53 mmol). Additional THF (158 ml) was added to flush celite. To the mixture was added (at -5 ° C) NMM (5.8 ml, 52.88 mmol) and after 5 minutes a chilled (-5 ° C) freshly prepared solution containing: A5 (31.96 g, 89.81 mmol) NMM (16 ml, 145 mmol) and DMF (120 ml). The reaction mixture was allowed to warm to rt. and stirred for 14 hours. The crude reaction mixture was filtered and the solvent was removed under reduced pressure. EtOAc (300 ml) was added to the remaining DMF solution and washed successively with aq. HCl (200 ml, 0.1 N) aq. NaHCO ₃ (200 ml, sat.) And rinsed with brine (300 ml). The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. The resulting material was applied onto silica (EtOAc as a solvent) and chromatographed on silica gel, eluting with EtOAc: hex solvent gradient. 1: 5 - 1: 1, which gives SAPLA4 (26.8 g, 78%) as a viscous colorless oil. The specified product is a 1: 1 mixture of diastereomers, Rf = 0.5 (silica, hex. / EtOAc 1: 1, dark blue / vanillin).

IR (film, DSM) 3295, 3060 and 3040, 2957, 2934, 2880, 2858, 1736, 1634, 1528, 1454, 1387, 1252, 1171, 1070 cm ^-1 .

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.01 (s, 3H), 0.03 (s, 3H), 0.85-0.97 (m, 12H), 0.92 (s, 9H ), 0.93 (s, 9H), 1.33 (d, J = 7.0, 3H), 1.37 (d, J = 7.0, 3H), 2.40-2.65 (m , 3H), 1.92-2.28 (m, 4H), 3.64-3.76 (m, 1H), 4.69-4.82 (m, 1H), 5.05 (d, J = 11.8, 1H), 5.20 (d, J = 11.8, 2H), 6.73 (d, J = 8.9, 1H), 6.98 (d, J = 9.0, 1H), 7.34 (br s, 5H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ -5.24, -4.81, 15.70, 17.43, 17.57, 18.84, 21.48, 21.61, 18.05, 23.28, 24.43, 24.55, 24.76, 25.68, 28.87, 31.36, 31.77, 41.27, 41.67, 48.68, 48.55, 48, 89, 58.71, 66.84, 83.84, 83.29, 128.09, 128.47, 135.40, 169.24, 170.67, 170.89, 171.16, 171.20, 209.11, 211.62.

m / z (FAB) 611.5 [(M + Na) ⁺ , 15], 589.5 [(M + H) ⁺ , 100]; m / z (FABHRMS) 589.369 045, C ₃₂ H ₅₂ N ₂ O ₆ Si requires (M + H) ⁺ , 589.367 291

Example 12

Synthesis of Boc-Aip-Leu-Pro-OBn (SNPLA4)

To a degassed solution of SNPLB2 (2.3 g, 6.32 mmol) in dry THF (30 ml) was added 10% Pd / C (0.74 g, 16 wt.%) And then hydrogenated at atmospheric pressure for 5 h 30 min or until complete conversion, according to TLC (disappearance of starting material). The resulting mixture was filtered through a pad of celite and additional THF (20 ml) was added to rinse the celite. To the filtered solution (at -5 ° C) BOP-Cl (1.77 g, 6.96 mmol) and NMM (765 μl, 6.91 mmol) were added and after 30 minutes a cooled (-5 ° C) solution containing A5 (3.15 g, 8.85 mmol), NMM (1.88 ml, 8.84 mmol) and DMF (14 ml) obtained 10 min before addition. The reaction mixture was allowed to warm to rt. and stirred for 17 hours. The crude reaction mixture was filtered and the solids washed with EtOAc (100 ml). The combined organic solutions were washed successively with aq. KHSO ₄ (50 ml, 10%), aq. NaHCO ₃ (50 ml, sat.) And brine (50 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo, and the resulting material was chromatographed on silica gel, eluting with a solvent gradient of EtOAc: hex. 1: 4 - 1: 1, which gives SNPLA4 (750 mg, 20%) as a white solid. The specified product is a mixture of diastereomers. Rf = 0.26 and 0.17 (silica, hex. / EtOAc 1: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.74-1.04 (m, 12H), 1.31-1.70 (m, 6H), 1.43 (s, 9H), 2.01 (m, 3H), 2.22 (m, 2H), 3.60 (m, 2H), 3.77 (m, 1H), 4.40 (m, 1H), 4.58 (m, 1H) 4.69 (m, 1H), 5.14 (m, 2H), 6.75 (d, J = 8.7, 1H), 7.04 (d, J = 7.8, 1H), 7 34 (br s, 5H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 16.74, 16.91, 20.22, 21.94, 23.53, 24.80, 25.06, 28.48, 29.14, 41, 29, 41.41, 46.98, 49.45, 49.64, 51.26, 59.06, 63.70, 64.27, 67.08, 79.86, 80.10, 128.32, 128.48, 128.74, 135.74, 156.28, 169.15, 169.32, 171.18, 171.91.

ESI-MC Calculate. for C ₃₁ H ₄₇ N ₃ O ₇ : 573.34. Found (m / z): 574.4 [(M + H)] ⁺ .

Example 13

Synthesis of Hiv-Leu-Pro-OBn (SHPLA4)

To a flask containing A5 (2.06 g, 4.78 mmol) in DCM (5 ml) at 0 ° C, NMM (506 mg, 5.01 mmol) was added with stirring. After 15 minutes, (2S) -2-hydroxy-3-methylbutanoic acid (hydroxyisovalerianic acid) (487 mg, 4.78 mmol) and DCC (986 mg, 4.78 mmol) were added portionwise. The reaction mixture was allowed to warm to 23 ° C and stirred for 14 hours. The suspension was diluted with CHCl ₃ (25 ml) and partitioned between aq. HCl (10 ml, 1n), aq. NaHCO ₃ (10 ml, sat.) And brine (10 ml), dried (Na ₂ SO ₄ ) and concentrated under reduced pressure. The residue was purified by flash LC (silica gel, hex. Gradient - EtOAc 1: 1 - 1: 3) to give SHPLA4 (1.13 g, 80%) as a white solid. Rf = 0.46 (hex. - EtOAc 1: 2).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.81, (d, J = 7.0, 3H) 0.92 (m, 6H), 0.97 (d, J = 7.0, 3H) 1.42 (m, 1H), 1.63 (m, 2H), 2.00 (m, 3H), 2.19 (m, 2H), 3.60 (m, 1H), 3.85 ( m, 1H), 3.88 (d, J = 4.8, 1H), 4.46 (m, 1H), 4.80 (m, 1H), 5.06 (d, J = 12.3, 1H), 5.14 (d, J = 12.3, 1H), 7.32 (m, 5H), 7.41 (d, J = 8.4, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 15.32, 19.08, 21.38, 23.11, 24.41, 24.69, 28.77, 31.31, 40.54, 46, 81, 48.20, 58.82, 66.69, 75.76, 127.93, 128.14, 128.37, 135.28, 171.39, 171.95, 173.94.

Example 14

Synthesis of Boc-Val-Leu-Pro-OBn (SVPLA4)

To a flask containing Boc-valine-OH (652 mg, 3 mmol) in DCM (6 ml) at 0 ° C., NMM (0.35 ml, 3.15 mmol) was added. After stirring for 15 minutes, A5 (1.065 g, 3 mmol), HOBt (405 mg, 3.0 mmol) and DCC (650 mg, 3.15 mmol) were added in portions. The reaction mixture was allowed to warm to 23 ° C and stirred for 14 hours. The suspension was diluted with DCM (25 ml) and washed successively with aq. KHSO ₄ (2 × 10 ml, 10%), aq. NaHCO ₃ (2 × 10 ml, sat.) And brine (10 ml), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash LC (silica, hex. Gradient - EtOAc 2: 1 - 1: 1) to give SVPIA4 (1.48 g, 93%) as a white solid. Rf = 0.57 (hex. - EtOAc 1: 2).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.83-0.95 (m, 12H), 1.2-1.4 (m, 1H), 1.42 (s, 9H), 1.40 -1.51 (m, 1H), 1.60-1.75 (m, 1H), 1.82-2.20 (m, 5H), 3.50-3.60 (m, 1H), 3 74-3.78 (m, 1H), 3.91 3.96 (m, 1H), 4.52-4.57 (s, 1H), 4.75-4.77 (m, 1H), 5.04 (br s, 1H), 5.05 (d, J = 12.3, 1H), 5.17 (d, J = 12.3, 1H), 6.60 (d, J = 8 4, 1H); 7.26-7.35 (m, 5H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 7.61, 19.19, 21.66, 23.23, 24.43, 24.78, 24.87, 25.55, 28.20, 28, 87, 30.91, 33.86, 41.68, 46.73, 48.86, 58.77, 59.74, 66.82, 79.66, 128.08, 128.21, 128.46, 135.47, 155.66, 170.83, 171.33, 171.60.

ESI-MC Calculate. for C ₂₈ H ₄₃ N ₃ O ₆ : 517.32. Found (m / z): 518.2 [(M + H)] ⁺ .

Example 15

Synthesis of Boc-Me-Val-Leu-Pro-OBu (SMPLA4)

Following the procedure developed for the synthesis of SVPIA4, starting from A5 (1.07 mg, 3.00 mmol) and Boc- (Me) Val-OH (694 mg, 3.00 mmol), the title compound was obtained as a white solid (1.39 g, 87%), after purification by flash LC (silica gel, hex. Gradient - EtOAc 2: 1 - 1: 1). Rf = 0.51 (hex. - EtOAc 1: 2).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.83-0.91 (m, 12H), 1.45 (s, 9H), 1.93-2.03 (m, 4H), 2.18 -2.22 (m, 2H), 2.76 (s, 3H), 3.40-3.50 (m, 2H) 3.55-3.62 (m, 1H), 3.75-3, 85 (m, 1H), 4.00-4.10 (m, 1H), 4.50-4.60 (m, 1H), 4.70-4.82 (m, 1H), 5.07 ( d, J = 11.2, 1H), 5.24 (d, J = 11.2, 1H), 6.20 (m, 0.5H), 6.50 (m, 0.5H), 7, 26-7.35 (m, 5H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 18.43, 19.77, 21.53, 23.23, 24.47, 24.77, 25.89, 25.30, 28.85, 41, 25, 46.67, 48.51, 58.75, 64.05, 66.79, 128.06, 128.18, 128.44, 135.49, 170.12, 170.80, 171.66.

ESI-MC Calculate. for C ₂₉ H ₄₅ N ₃ O ₆ : 531.33. Found (m / z): 532.3 (M + H) ⁺ .

Example 16

Hip-Leu-Pro-OBn Synthesis (SAPLA3)

In a flask containing SAPLA4 (15.86 g, 26.97 mmol), a clear, colorless solution of 1M tetrabutylammonium fluoride in THF (80.9 ml, 80.9 mmol) was added and the mixture was vigorously stirred at rt. 15 min (or until complete conversion by TLC). The reaction was quenched by addition of H ₂ O (4 ml) and silica gel (50 g). The crude material was concentrated and purified by flash LC (silica gel, hex.: EtOAc 2: 1 - 1: 1) to give SAPLA3 (12.2 g, 95%) as a white solid (mixture of diastereomers). Rf = 0.36 and 0.29 (silica, hex.: CHCl ₃ : IPA; 1: 5: 1).

IR (film, DCM) ν 3450-3293, 3060 and 3040, 2961, 2946, 2883, 2852, 1746, 1632, 1533, 1454, 1357, 1387, 1265, 1173, 1095, 1045, 1018 cm ^-1 .

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.71 (d, J = 6.8, 3H), 0.81 (d, J = 6.6, 3H), 0.88 (d, J = 6.5, 3H), 0.91 (d, J = 6.5, 3H), 0.94 (d, J = 6.5, 3H), 0.99 (d, J = 7.1, 3H ), 1.07 (d, J = 6.5, 3H), 1.36 (d, 6.5, 3H), 1.43-1.52 (m, 2H), 1.60-1.66 (m, 1H), 1.93-2.10 (m, 3H), 2.12-2.23 (m, 2H), 3.53-3.58 (m, 1H), 3.65 (q , J = 7.1, 1H), 3.67-3.73 (m, 1H), 3.89 (q, J = 7.1, 1H), 3.96 (d, J = 4.2, 1H), 4.22 (d, J = 4.1, 1H), 4.54-4.56 (m, 1H), 4.58-4.62 (m, 1), 4.69-4, 73 (m, 1H), 5.1 (d, J = 12.1, 1H), 5.18 (d, J = 12.1, 1H), 6.57 (d, J = 8.5, 1H ), 6.63 (d, J = 8.5, 1H), 7.28-7.38 (m, 5H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 14.06, 14.26, 15.85, 16.48, 20.07, 20.53, 22.02, 22.25, 25.37, 25, 46, 29.45, 29.53, 31.59, 32.09, 41.13, 42.29, 49.93, 50.91, 51.02, 59.52, 67.60, 81.02, 128.78, 1.29.2, 169.48, 171.58, 172.17, 209.76.

m / z (FAB) 497.4 [(M + Na) ⁺ , 12], 475.5 [(M + H) ⁺ , 100]; m / z (FABHRMS) 497.263 162, C ₂₆ H ₃₈ N ₂ O ₆ requires (M + Na) ⁺ , 497.262 757. Anal. Calculate. for C ₂₆ H ₃₈ N ₂ O ₆ : C, 65.82; H, 8.02; N, 5.91. Found: C, 65.97; H, 8.18; N, 5.76.

Example 17

Aip-Leu-Pro-OBn Synthesis (SNPLA3)

To a solution of SNPLA4 (750 mg, 1.30 mmol) in dioxane (15 ml, anhydrous) was added a solution of hydrogen chloride in dioxane (39 ml, 5.3 n) and the mixture was stirred for 5 h or until complete conversion by TLC (disappearance of the original material). The solution was concentrated under reduced pressure and the resulting oil was distilled with CHCl ₃ (15 ml), MTBE (15 ml) and hex. (15 ml). The residue was dried in vacuo to remove residual HCl to give a foamy solid. SNPLA3 (660 mg, quant.) Is used directly in the next step without further purification.

ESI-MC Calculate. for C ₂₆ H ₃₉ N ₃ O ₅ : 473.29. Found 474.2 (M + H) ⁺ .

Example 18

Synthesis of Val-Leu-Pro-OBn (SVPLA3)

In a flask containing SVPLA4 (215 mg, 0.41 mmol), a solution of hydrogen chloride in dioxane (1.5 ml, 5.3 N) was added and the mixture was stirred for 5 h or until complete conversion by TLC. The solution was concentrated under reduced pressure and the resulting oil was distilled with CHCl ₃ (5 ml), MTBE (5 ml) and hex. (5 ml). The residue was dried in vacuo to remove residual HCl, a foamy solid was obtained, SVPLA3 (185 mg, quant.) Was used directly in the next step without further purification.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.86-0.90 (m, 6H), 1.04 (d, J = 6.3, 3H), 1.12 (d, J = 6, 3, 3H), 1.45-1.55 (m, 1H), 1-60-1.80 (m, 2H), 1.82-2.11 (m, 2H), 2.11-2, 25 (m, 1H), 2.25-2.40 (m, 1H), 3.50-3.70 (m, 1H), 3.80-3.95 (m, 2H), 4.52- 4.57 (s, 1H), 4.70-4.85 (m, 1H), 5.05 (d, J = 12, 1H), 5.20 (d, J = 12.3, 1H), 7.27-7.37 (m, 5H), 7.91 (m, 1H), 8.62 (br s, 2H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 18.57, 18.79, 21.83, 23.11, 24.50, 24.67, 24.90, 25.34, 28.92, 30, 23, 33.25, 40.40, 47.04, 49.46, 49.94, 59.26, 60.02, 66.88, 128.16, 128.27, 128.51, 135.48, 167.54, 170.80, 171.94.

Example 19

Synthesis (Me) Val-Leu-Pro-OBn (SMPLA3)

Following the procedure developed for the synthesis of SVPLA3, starting from SMPLA4 (940 mg, 1.94 mmol), the title compound (828 mg, quant.) Was obtained as a white solid. The specified product is used directly in the next step without further purification.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.93 (d, J = 6.3, 6H), 1.07 (d, J = 6.3, 3H), 1.21 (d, J = 6.3, 3H), 1.47 (m, 1H), 1.73 (m, 2H), 2.00 (m, 3H), 2.23 (m, 1H), 2.52 (m, 1H ), 2.83 (br s, 3H), 3.56-3.65 (m, 2H), 3.77 (m, 1H), 4.59 (m, 1H), 4.66 (m, 1H), 5.07 (d, J = 12.3, 1H), 5.19 (d, J = 12.3, 1H), 7.27-7.38 (m, 5H), 7.90 ( m, 1H), 9.11 (m, 0.5H), 9.61 (m, 0.5H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 13.98, 18.37, 19.57, 21.33, 22.52, 23.16, 24.74, 28.77, 29.78, 31, 54, 32.54, 39.87, 46.75, 50.09, 58.91, 66.85, 122.12, 128.06, 128.24, 128.47, 135.43, 166.22, 170.73, 171.54.

ESI-MC Calculate. for C ₂₄ H ₃₈ CIN ₃ O ₄ : 431.2. Found (m / z): 432.3 (M + H) ⁺ .

Example 20

Boc-Ist Synthesis (TBDMS) -Hip-Leu-Pro-OBn (SAPLA2)

To a solution of SAPIA3 (12.2 g, 25.44 mmol) in bezv. DCM (75 ml) at -5 ° C in an Ar atmosphere was added in portions of DMAP (0.932 g, 7.6 mmol), C1 (11.89 g, 30.53 mmol) and DCC (6.613 g, 32.05 mmol), withstanding a temperature of <-5 ° C (ice-salt bath). The reaction mixture was stirred for 14 hours at -5 ° C and then filtered and concentrated. The crude material is distilled with ACN, cooled (-10 ° C), filtered and concentrated again. The resulting product was dissolved in EtOAc (400 ml) and washed successively with aq. KHSO ₄ (2 × 200 ml, 10%), brine (200 ml), aq. NaHCO ₃ (200 ml, sat.) And rinse with brine (200 mL). The organic phase was dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give a colorless oil, which was chromatographed on silica gel, eluting with hex solvent. - EtOAc 3: 1 - 2: 1, which gives SAPLA2 (19.35 g, 90%) as a white foam (mixture of diastereomers).

IR (film, DCM) ν 3365-3200, 3069, 3038, 2959, 2930, 2882, 2857, 1746, 1688, 1640, 1533, 1456, 1389, 1258, 1171, 1086 cm ^-1 .

¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.01 (s, 3H), 0.03 (s, 3H), 0.05 (s, 3H), 0.07 (s, 3H), 0, 77-1.03 (m, 18H), 0.84 (s, 9H), 0.85 (s, 9H), 1.33 (d, J = 7.4, 3H), 1.32-1, 36 (m, 2H), 1.49 (d, J = 7.5, 3H), 1.38-1.62 (m, 3H), 1.42 (s, 9H), 1.44 (s, 9H), 1.51-1.77 (m, 1H), 1.88-2.37 (m, 3H), 2.17-2.33 (m, 2H), 2.47-2.74 ( m, 2H), 3.34-3.72 (m, 1H), 3.72-3.82 (m, 1H), 3.99-4.40 (m, 1H), 4.03-4, 16 (m, 1H), 4.49 (d, J = 10.3, 1H), 4.54-4.59 (m, 1H), 4.63-4.70 (m, 2H), 4, 75 (d, J = 4.5, 1H), 4.77-4.81 (m, 1H), 4.95-5.19 (m, 2H), 5.22 (d, J = 5.2 , 1H), 5.32 (d, J = 10.5, 1H), 6.38 (d, J = 10.9, 1H), 6.71 (d, J = 7.4, 1H), 6 76 (d, J = 8.4, 1H); 8.60 (d, J = 9.5, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ -5.05, -4.49, 11.83, 12.03, 13.01, 13.51, 13.83, 14.08, 16.92, 17.10, 17.85, 19.14, 19.65, 21.57, 22.09, 22.96, 23.28, 24.36, 24.60, 24.85, 25.73, 26, 97, 27.33, 28.35, 28.46, 28.93, 29.09, 29.65, 34.12, 34.16, 40.45, 40.85, 41.18, 42.20, 46.74, 46.16, 47.99, 48.34, 48.90, 49.42, 57.62, 58.81, 58.96, 60.46, 66.62, 66.88, 68, 18, 69.69, 78.98, 79.24, 79.84, 82.95, 128.08-128.49, 135.48 135.61, 155.85, 158.27, 157.44, 168 40, 169.07, 170.65, 170.86, 171.42, 171.79, 203.09 205.97.

m / z (FAB) 846.6 [(M + H) ⁺ , 15], 746.6 (100); m / z (FABHRMS) 868.516 630, C ₄₆ H ₇₅ N ₃ O ₁₀ Si requires (M + Na) ⁺ 868.511 930

Example 21

Synthesis of Boc-Ist (TBDMS) -Aip-Leu-Pro-OBn (SNPLA2)

NMM (0.19 ml) was added to a flask containing SNPLA3 (hydrochloride) (660 mg, 1.12 mmol) in DCM (15 ml, anhydrous) at 0 ° C. After 15 minutes, C1 (632 mg, 1.62 mmol), HOBt (266 mg, 1.73 mmol) and DCC (331 mg, 1.60 mmol) were added in portions. The flask was allowed to warm to room temperature and stirring was continued overnight. The crude reaction mixture was partitioned between DCM (50 ml) and aq. KHSO ₄ (2 × 20 ml, 10%). The organic phase is washed successively with aq. NaHCO ₃ (2 × 20 ml, sat.) And brine (20 ml), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The resulting white solid was purified by flash LC (silica gel, hex. Gradient - EtOAc 4: 1 - 1: 1) to give the title compound as a white solid (700 mg, 73%, diastereomer mixture).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.05 (s, 3H), 0.08 (s, 3H), 0.09 (s, 3H), 0.68-1.05 (18H, m ), 0.87 (9H, s), 1.05-1.85 (12H, m), 1.42 (9H, s), 1.85-2.10 (3H, m), 2.11- 2.31 (2H, m), 2.32-2.46 (2H, m), 2.47-2.60 (m, 2H), 3.34-3.90 (2H, m), 3, 93-4.30 (2H, m), 4.50-4.89 (6H, m), 4.90-5.12 (2H, m), 5.07 (d, J = 12.2, 2H ), 5.18 (d, J-12.2, 2H), 5.60 (1H, d, J = 9.7), 5.67 (1H, d, J = 10.2), 5.89 (1H, d, J = 11.2), 6.56 (1H, d, J = 7.3), 6.70 (1H, d, J = 8.3), 6.76 (1H, d, J = 6.8), 6.94 (d, J = 6.8, 1H), 7.01-7.19 (m, 1H), 7.32 (br s, 5H), 8.17 ( 1H, d, J = 7.8), 8.28 (d, J = 7.8, 1H).

ESI-MC Calculate. for C ₄₅ H ₇₉ N ₄ O ₉ Si: 844.54. Found (m / z): 845.5 (M + H) ⁺ .

Example 22

Synthesis of Boc-Ist (TBDMS) -Hiv-Leu-Pro-OBn (SHPLA2)

Following the procedure developed for the synthesis of SAPLA2, based on SHPLA4 (850 mg, 2.0 mmol) and C1 (935 mg, 2.4 mmol), the title compound was obtained (1.53 g, 97%) after purification by flash LC ( silica gel, hex gradient - EtOAc 3: 1 - 2: 1). Rf = 0.63 (hex. - EtOAc 2: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) mixture of BocNH rotamers: δ 0.04 (s, 3H), 0.06 (s, 3H), 0.88 (s, 9H), 0.78-1.04 (m, 18H), 1.10-2.80 (m, 11H), 1.44 (s, 9H), 1.46 (s, 9H), 3.57 (m, 2H), 3.74 ( m, 1H), 3.85 (m, 1H), 4.03 (m, 1H), 4.23 (d, J = 4.8, 1H), 4.48 (m, 1H), 4.85 (m, 1H), 4.90 (d, J = 10, 1H), 5.05 (m, 1H), 5.20 (d, J = 10, 1H), 5.23 (d, J = 10 , 1H), 6.64 (d, J = 6.4, 1H), 6.88 (d, J = 8.6, 1H), 7.32 (m, 5H), 8.54 (d, J = 8.3, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ -5.14, -4.58, 11.84, 12.97, 17.78, 17.92, 18.98, 21.05, 23.11, 23.49, 25.61, 26.92, 28.36, 28.70, 30.12, 33.68, 38.72, 42.86, 46.51, 48.18, 58.67, 60, 24, 66.52, 71.14, 79.40, 82.66, 127.96, 128.01, 128.33, 135.36, 157.30, 169.92, 171.10, 171.69, 171.97.

ESI-MC: Calculate. for C ₄₂ H ₇₁ N ₃ O ₉ : 789.50. Found 790.5 (M + H) ⁺ .

Example 23

Synthesis of Boc-Ist (TBDMS) -VaI-Leu-Pro-OBn (SVPLA2)

Follow the procedure developed for the synthesis of SNPLA2 based on SVPLA3 (hydrochloride) (1.2 g, 2.64 mmol), C1 (1.23 g, 3.17 mmol), DCC (654 mg, 3.17 mmol), HOBt (464 mg, 3.43 mmol), NMM (0.35 ml) and DCM (6 ml). The title compound was obtained as a white solid (1.87 g, 89%) after purification by flash LC (silica gel, hex. Gradient - EtOAc 3: 1 - 2: 1).

¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.07 (br s, 6H), 0.81-0.96 (m, 27H), 1.11-1.38 (m, 3H), 1 , 39-1.47 (br.s, 7H), 1.51 (br.s, 3H), 1.58-1.70 (m, 3H), 1.70-1.84 (m, 1H) 1.86-2.60 (m, 4H), 2.28-2.58 (m, 2H), 3.58-3.62 (m, 1H), 3.62-3.73 (m, 1H), 3.73-3.90 (m, 1H), 4.05-4.12 (m, 1H), 4.13-4.19 (m, 1H), 4.19-4.23 ( m, 1H), 4.49-4.54 (m, 1H), 4.77-5.06 (m, 2H), 5.18 (d, J = 12.3, 1H), 5.55 ( broad s, 1H), 6.44-6.61 (m, 2H), 7.30-7.35 (m, 5H), 7.94-7.98 (m, 1H).

ESI-MC Calculate. for C ₄₂ H ₇₂ N ₄ O ₈ Si: 788.51. Found (m / z): 789.5 (M + H) ⁺ .

Example 24

Boc-Ist Synthesis (TBDMS) - (Me) Val-Leu-Pro-OBn (SMPLA2)

To a flask containing SMPLA3 (hydrochloride) (176 mg, 0.38 mmol) in DCM (2 ml, anhydrous) at 0 ° C was added NMM (41 μl, 0.38 mmol). After 15 minutes, C1 (176 mg, 0.46 mmol) and DCC (93 mg, 0.46 mmol) were added in portions. The flask was allowed to warm to room temperature and stirring was continued overnight. The reaction mixture was partitioned between DCM (10 ml) and aq. KHSO ₄ (2 × 5 ml, 10%). The organic phase is washed successively with aq. NaHCO ₃ (2 × 5 ml, sat.) And brine (5 ml), dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The resulting white solid was purified by flash LC (silica, gradient hex. EtOAc 3: 1 - 2: 1) to give SMPLA2 (127 mg, 42%) as a white solid. Rf = 0.51 (hex. - EtOAc 1: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) δ -0.08 (s, 3H), 0.05 (s, 3H), 0.75-0.86 (m, 27H), 1.00-1, 43 (m, 11H), 1.52-1.65 (m, 1H), 1.68-1.80 (m, 1H), 1.83-2.01 (m, 3H), 2.08- 2.24 (m, 2H), 2.40 (m, 2H), 2.87 (s, 3H), 3.50-3.57 (m, 3H), 3.71-3.76 (m, 1H), 4.29 (m, 1H), 4.47-4.63 (m, 4H), 5.01 (d, J = 12.9, 1H), 5.11 (d, J = 12, 9, 1H), 6.41 (d, J = 7.8, 0.5H), 6.62 (d, J = 7.8, 1H), 7.01 (d, J = 7.8, 0 5H); 7.23-7.28 (m, 5H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.56, 13.57, 13.98, 17.83, 18.87, 19.46, 21.77, 22.94, 23.11, 24, 50, 24.71, 24.80, 25.64, 25.76, 25.97, 27.29, 28.18, 28.75, 30.40, 34.18, 39.28, 40.85, 46.58, 48.61, 57.03, 58.65, 62.26, 66.63, 69.21, 78.72, 127.93, 128.05, 128.11, 128.33, 135, 42, 155.91, 169.76, 170.48, 171.56, 172.01.

ESI-MC Calculate. for C ₄₃ H ₇₄ N ₄ O ₈ Si: 802.53. Found (m / z): 825.5 (M + Na) ⁺ .

Example 25

Synthesis of Ist-Hip-Leu-Pro-OBn (SAPLA1)

To a solution containing SAPLA2 (19.32 g, 22.8 mmol) in bezv. dioxane (78 ml), add a solution of hydrochloric acid in anhydrous dioxane (4.2 n, 220 ml, 924 mmol). The resulting solution was stirred at 21 ° C for 4.30 hours or until the starting material (TLC) completely disappeared. Then, the solution was concentrated under reduced pressure. The residue was dissolved in DCM (25 ml) and concentrated to remove residual HCl. The resulting residue was dried in vacuo to completely remove traces of free HCl (3 h), yielding 17.3 g of SAPLA1 (15.1 g, quant.) As a white foam (mixture of diastereomers).

Example 26

Synthesis of Ist-Aip-Leu-Pro-OBn (SNPLA1)

Following the procedure developed for the synthesis of SAPLA1, starting from SNPLA2 (700 mg, 0.82 mmol), the title compound is obtained as a white solid (545 mg, quant.), After precipitation of Et ₂ O (mixture of diastereomers).

¹ H-NMR (300 MHz, CDCl ₃₎ δ 0,86-1,04 (m, 18H), 1,02-1,22 (m, 3H), 1,23-1,58 (m, 5H) 1.60-1.80 (m, 2H), 1.82-2.01 (m, 3H), 2.24 (m, 2H), 2.40-2.85 (m, 2H), 3 24 (m, 1H), 3.45 (m, 1H), 3.60 (m, 1H), 3.70-4.05 (m, 2H), 4.46 (m, 2H), 4, 47-4.75 (m, 2H), 5.10 (br.s, 2H), 7.34 (br.s, 5H), 7.98 (br.s, 1H), 8.10 (br. s, 1H).

ESI-MC Calculate. for C ₃₄ H ₅₄ N ₄ O ₇ : 630.40. Found (m / z): 631.4 (M + H) ⁺ .

Example 27

Synthesis of Ist-Hiv-Leu-Pro-OBn (SHPLA1)

Following the procedure developed for the synthesis of SAPLA1, starting from SHPLA2 (1.53 g, 1.94 mmol), the title compound (1.12 g, quant.) Is obtained as a white solid after precipitation of Et ₂ O.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.86-1.04 (m, 18H), 1.10-1.22 (m, 3H), 1.42 (m, 2H), 1.70 (m, 2H), 1.97 (m, 3H), 2.24 (m, 2H), 2.83 (m, 1H), 2.97 (m, 1H), 3.34 (m, 1H) 3.61 (m, 1H), 3.75 (m, 1H), 3.90 (m, 1H), 4.56 (m, 2H), 4.75 (m, 1H), 5.04 ( d, J = 11, 1H), 5.18 (d, J = 11, 1H), 7.34 (br s, 5H), 8.21 (br s, 3H).

ESI-MC Calculate. for C ₃₁ H ₄₉ N ₃ O ₇ : 575.36. Found (m / z): 576.3. (M + H) ⁺ .

Example 28

Synthesis of Ist-Val-Leu-Pro-OBn (SVPLA1)

Following the procedure developed for the synthesis of SAPLA1, starting from SVPLA2 (1.87 g, 2.36 mmol), the title compound (1.40 g, quant.) Is obtained as a white solid after precipitation of Et ₂ O.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.01 (m, 15H), 1.01-1.10 (m, 3H), 1.20-1.79 (m, 5H) 1.81-2.05 (m, 4H), 2.05-2.15 (m, 2H), 2.50-2.68 (m, 2H), 2.82-3.1 (m, 1H), 3.20-3.35 (m, 1H), 3.50-3.70 (m, 1H), 3.80-3.90 (m, 1H), 4.18-4.30 ( m, 1H), 4.35-4.45 (m, 1H), 4.45-4.55 (m, 1H), 4.60-4.70 (m, 1H), 5.02 (d, J = 12.3, 1H), 5.15 (d, J = 12.3, 1H), 7.28-7.38 (m, 5H), 7.5 (br.s, 1H), 7, 9 (br.s, 3H), 8.15 (br.s, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 10.81, 14.67, 18.29, 19.32, 21.48, 23.14, 24.48, 24.76, 25.81, 30, 29, 33.41, 40.31, 46.96, 49.35, 59.14, 59.94, 60.67, 66.94, 128.11, 128.32, 128.54, 135.32, 171.58, 171.74, 171.80, 172.57.

ESI-MC Calculate. for C ₃₁ H ₅₀ N ₄ O ₆ : 574.35. Found (m / z): 575.3 (M + H) ⁺ .

Example 29

N-tert-butyloxycarbonyltreonine phenacyl ester (D3)

To a stirred suspension of Boc-Thr-OH (21.91 g, 0.1 mol) in EtOAc (200 ml) at 0 ° C was added TEA (14 ml, 0.1 mol) and bromoacetophenone (19.0 g, 0, 1 mol). The reaction mixture was allowed to warm to 20 ° C, stirred for 2 days and then diluted with EtOAc (500 ml). After washing, successively aq. HCl (200 ml, 0.1 n), H ₂ O (100 ml), aq. NaHCO ₃ (200 ml, 1 N) and brine (200 ml) and drying (Na ₂ SO ₄ ), filtered and concentrated in vacuo, the residue was triturated with Et ₂ O and filtered. The resulting solid was dried in the dark to give D3 (28.6 g, 85%). Rf = 0.55 (hex. - EtOAc 1: 1, silicon dioxide); mp = 114.2 ° C; [α] _D ²² -29.4 (c 2, EtOAc).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.31 (d, J = 6.6, 3H), 1.46 (s, 9H), 3.77 (brd, OH), 4.44 (dd, J = 9.6, 1H), 4.6 (q, 1H), 5.34 (d, J = 16.5, 1H), 5.37 (brd, OH) 5.68 ( d, J = 16.8, 1H), 7.51 (t, 2H), 7.65 (t, 1H), 7.92 (dd, 2H).

Example 30

N-Benzyloxycarbonyl-N, O-dimethyl-L-tyrosine (E1)

To a stirred solution of Z-Tyr-OH (63.24 g, 200 mmol) in THF (900 ml) at 0 ° C, finely ground KOH (112.72 g, 2 mol) was added in portions, followed by tetrabutylammonium bisulfate (6.36 g , 10 wt.%). Then, dimethyl sulfate (127.2 ml, 1.33 mol) was added dropwise over 30 minutes while maintaining the reaction mixture at a temperature below 4 ° C. The reaction mixture was stirred an additional 30 minutes and H ₂ O (950 ml) was added. After stirring for 5 hours at 0 ° C., the reaction mixture was diluted with diethyl ether (1500 ml), the aqueous layer was separated and the organic layer was extracted with aq. NaHCO ₃ (2 × 500 ml, sat.). The combined aqueous layers were acidified to pH 1 with aq. 1M KHSO ₄ and extracted with EtOAc (5 × 500 ml). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was precipitated with diethyl ether and filtered to give E1 as a white solid (53.85 g, 78%).

[α] _D ²² -57.16 (c 2.23 CHCl ₃ ) (lit. [α] _D -48 (c = 2.23 CHCl ₃ ) JACS, 112, 21, 1990).

Example 31

O- (benzyloxycarbonyl-N, O-dimethyl-L-tyrosyl) N-tert-butyloxycarbonyl-L-threonine phenylacyl ester (D2)

To a solution of D3 (33.72 g, 100 mmol) in DCM at 0 ° C was added DMAP (3.66 g, 30 mmol) and E1 (34.33 g, 100 mmol). After stirring for 10 min at 0 ° C, DCC (22.7 g, 110 mmol) was added. The reaction mixture was allowed to warm to room temperature and was stirred overnight. The mixture was then filtered and the filtrate was concentrated to dryness. The residue was distilled with ACN (100 ml), filtered again and the filtrate was concentrated. The residue was dissolved in EtOAc (200 ml) and partitioned sequentially between aq. KHSO ₄ (100 ml, 10%), aq. NaHCO ₃ (100 ml, sat.) And brine (100 ml). The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was purified by flash LC (silica gel, EtOAc-hex gradient 1: 4 - 1: 2) to give D2 (65.5 g, 98%). [α] _D ²² -39.56 (c 1,06 CHCl _3); Rf = 0.55 (EtOAc: hex. 1: 1).

Example 32

O- (Benzyloxycarbonyl-N, O-dimethyl-L-tyrosyl) -N-tert-butyloxycarbonyl-L-threonine (D1)

To a homogeneous solution of D2 (24.49 g, 38.4 mmol) in aq. AcOH (211 ml, 90%), Zn powder (18.65 g, 288.3 mmol) was added at 0 ° C. The resulting mixture was stirred at 0 ° C for 3 hours until the starting material disappeared (by TLC). The reaction mixture was filtered through celite and washed with EtOAc (200 ml). The filtrate was concentrated under reduced pressure and the residue was distilled with Et ₂ O (200 ml) and filtered. The filtrate was partitioned sequentially between aq. KHSO ₄ (100 ml, 10%) and brine (100 ml). The organic phase was dried (Na ₂ SO ₄ ) and concentrated to give an oil which was purified by flash LC (Lichroprep RPC18, ACN: H ₂ O 1: 1 (800 ml, then 7: 3 (600 ml)] to give D1 (15, 53 g, 74%) as a white solid. [Α] _D ²⁴ -27.6 (s 2.187, DCM); lit. [α] _D -20.5 (s 2, DCM), JOC, 62, 2 1997. Rf = 0.58 [ACN / H ₂ O (7: 3)].

IR (film, DCM) ν 3400, 3050, 2900, 1715, 1613, 1514, 1456, 1402, 1368, 1248, 1165, 1061, 1036 cm ⁻¹ .

¹ H-NMR (200 MHz, CDCl ₃ ) δ 1.29 (d, J = 6.5, 3H), 1.45 (s, 9H), 2.74 (s, 3H), 2.75 (s , 3H), 2.76-3.31 (m, 2H), 3.77 (s, 3H), 4.42-4.52 (m, 1H), 4.66-4.83 (m, 1H ), 5.01-5.16 (m, 2H), 5.30-5.53 (m, 2H), 6.72-6.81 (m, 2H), 6.95-7.09 (m , 2H); 7.35 (br s, 5H).

¹³ C-NMR (75 MHz, CDCl ₃₎ δ 16.44, 16.82, 28.23, 31.71, 31.97, 33.82, 33.68, 55.14, 56.87, 56, 75, 60.39, 60.58, 67.51, 67.76, 71.83, 72.47, 80.40, 113.91, 127.59, 128.69, 129.77, 136.42, 156.00, 156.19, 156.71, 158.31, 159.47, 169.78.

m / z (FAB) 567,1 [ (M + Na) +, 46], 545,1 [(M + H ) +, 7], 445.1 (100); m / z (FABHRMS) 567.233 280, C ₂₈ H ₃₅ N ₂ O ₉ requires (M + Na) ⁺ 567.231 851.

Example 33

Synthesis of Boc-Thr (Z-N (Me) -O (Me) -Tyr) -Ist-Hip-Leu-Pro-OBn (SAPL7)

Into a flask containing HBTU (9.079 g, 23.9 mmol), HOBt (3.490 g, 22.8 mmol), SAPLA1 (15.258 g, 22.8 mmol) and D1 (12.417 g, 22.8 mmol) are introduced via cannula in Ar atmosphere at -5 ° С solution without. DCM (296 ml) and anv. DMF (148 ml). After 5 minutes of stirring, a DIPEA syringe (15.9 ml, 91.2 mmol) was added dropwise with a temperature below -5 ° C. The resulting reaction mixture was stirred for 21 hours at -5 ° C. MTBE (300 ml) and KHSO ₄ (200 ml, 10%) are added and the resulting mixture is filtered and concentrated to 300 ml. An additional amount of MTBE (200 ml) was added, the layers were separated, and the organic phase was successively treated with aq. KHSO ₄ (200 ml, 10%), brine (200 ml), aq. NaHSO ₄ (200 ml, sat.) And rinsed with brine (200 ml). The organic phase was dried (Na ₂ SO ₄ ) and concentrated under reduced pressure to give a yellow oil (30 g). The oil is dissolved in MTBE and treated with hex. with stirring. A solid precipitates, additional hex is added. The solid product was filtered to give SAPL7 (18.33 g, 69% yield) as a white solid. The specified product is a mixture of two diastereomers. Rf = 0.80 and 0.59 (hex.: EtOAc 1: 2).

IR (film, DCM) ν 3350, 2961, 2927, 2893, 1744, 1688, 1638, 1514, 1454, 1368, 1304, 1248, 1171, 1067, 1036 cm ^-1 .

¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.74-0.92 (m, 18H), 1.05-1.15 (m, 2H), 1.18-1.20 (m, 2H) 1.23 (d, J = 6.8, 3H), 1.25 (d, J = 6.8, 3H), 1.29 (d, J = 6.9, 3H), 1.42 ( s, 9H), 1.45 (s, 9H), 1.50-1.66 (m, 3H), 1.89-2.02 (m, 4H), 2.17-2.25 (m, 2H), 2.37-2.42 (m, 1H), 2.81 (s, 3H), 2.88 (s, 3H), 2.91 (s, 3H), 2.95 (s, 3H) ), 2.84-2.93 (m, 2H), 3.17-3.25 (m, 1H), 3.53-3.59 (m, 1H), 3.75 (s, 3H), 3.88-3.98 (m, 4H), 4.49 (d, J = 3.1, 1H), 4.51 (d, J = 3.1, 1H), 4.53-4.57 (m, 1H), 4.68-4.72 (m, 1H), 4.96-4.99 (m, 1H), 5.02-5.33 (m, 4H), 5.02 (d , J = 3.2, 1H), 5.23 (d, J = 3.1, 1H), 5.26-5.33 (m, 1H), 5.47 (1d, J = 9.5, 1H), 6.74 (d, J = 7.8, 2H), 6.77 (d, J = 7.7, 2H), 7.08 (d, J = 7.7, 2H), 7, 17 (d, J = 7.5, 1H), 7.21 (d, J = 9.5, 1H), 7.23-7.36 (m, 10H), 7.75 (d, J = 7 9, 1H); 7.79 (d, J = 8.2, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.95, 13.27, 15.16, 16.47, 17.33, 18.79, 21.28, 23.65, 24.65, 24, 72, 27.09, 28.08, 28.93, 31.20, 31.32, 33.62, 33.98, 38.38, 41.01, 47.12, 49.38, 54.96, 55.17, 57.89, 58.83, 60.01, 60.16, 67.18, 71.05, 71.32, 80.34, 81.24, 113.89, 127.51, 128, 59, 129.69, 129.77, 135.52, 136.77, 156.93, 158.27, 169.87, 170.62, 171.15, 171.85, 172.39, 204.88.

m / z (FAB) 1181.2 [(M + Na) ⁺ , 20], 1159.2 [(M + H) ⁺ , 80], 1059.2 (100). Anal calculated C ₆₂ H ₈₇ N ₅ O ₁₆ : C, 64.30; H, 7.502; N, 6.05. Found: C, 64.14; H, 7.66; N, 5.95.

Example 34

Synthesis of Boc-Thr (Z-N (Me) -O (Me) -Tyr) -Ist-Aip-Leu-Pro-OBn (SNPL7)

Following the procedure developed for the synthesis of SAPL7, starting from SNPLA1 (150 mg, 0.22 mmol) and D1 (122 mg, 0.22 mmol), the title compound was obtained as a white solid (130 mg, 51%), after purification by flash LC (silica gel, hex. gradient - EtOAc 2: 1 - 1: 3) (mixture of diastereomers).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.74-1.03 (m, 18H), 1.16-1.37 (m, 10H), 1.45 (s, 9H), 1.68 (m, 3H), 1.99 (m, 4H), 2.22 (m, 2H), 2.48 (m, 1H), 2.82 (s, 3H), 2.84-3.10 ( m, 2H), 3.19 (m, 1H), 3.51-3.69 (m, 2H), 3.75 (s, 3H), 3.72-4.02 (m, 3H), 4 18 (m, 1H), 4.50-4.73 (m, 4H), 5.00-5.27 (m, 5H), 5.49 (m, 2H), 6.54 (d, J = 9.2, 1H), 6.78 (d, J = 6.8, 2H), 7.02 (d, J = 6.8, 2H), 7.18 (m, 1H), 7.23 -7.36 (m, 10H); 7.52 (d, J = 6.8, 1H).

ESI-MC Calculate. for C ₆₂ H ₈₈ N ₆ O ₁₅ : 1156.63. Found 1158.3 (M + H) ⁺ .

Example 35

Synthesis of Boc-Thr (Z-N (Me) -O (Me) -Tyr) -Ist-Hiv-Leu-PrO-OBn (SHPL7)

Following the procedure developed for the synthesis of SAPL7, starting from SHPLA1 (1.12 g, 1.94 mmol) and SAPLD1 (544.6 mg, 1.94 mmol), the title compound (1.045 g, 61%) was obtained as white solid after purification by flash LC (silica gel, gradient hex. - EtOAc 1: 1 - 1: 2). Rf = 0.46 (hex. - EtOAc 1: 2).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.74-1.02 (m, 18H), 1.20 (m, 5H), 1.40 (m, 3H), 1.46 (s, 9H ), 1.62 (m, 2H), 1.82-2.20 (m, 3H), 2.20 (m, 2H), 2.50 (m, 1H), 2.78 (s, 3H) , 2.90 (m, 1H), 3.20 (m, 1H), 3.58 (m, 1H), 3.67 (s, 3H), 3.79 (m, 1H), 3.88 ( m, 1H), 4.06 (m, 2H), 4.40 (m, 2H), 4.82 (m, 2H), 4.94 (m, 1H), 4.98 (m, 1H), 5.08 (m, 1H), 5.28 (m, 3H), 5.56 (d, J = 6.2, 1H), 6.84 (d, J = 8.3, 2H), 6, 98 (d, J = 6.5, 1H), 7.07 (d, J = 8.3, 2H), 7.34 (m, 10H), 7.52 (d, J = 6.2, 1H )

¹³ C-NMR (75 MHz, CDCl ₃₎ δ 11.27, 13.77, 13.95, 16.75, 17.46, 18.31, 20.78, 20.96, 23.09, 24, 35, 24.53, 26.74, 27.93, 28.63, 30.17, 33.76, 39.06, 40.01, 46.69, 48.17, 54.89, 57.04, 57.68, 58.65, 60.20, 60.60, 66.74, 67.08, 68.31, 70.30, 78.49, 79.90, 113.62, 127.36, 127, 70, 128.14, 128.18, 128.31, 129.64, 135.19, 136.31, 155.86, 156.61, 158.09, 169.77, 170.70, 171.06, 171.17, 173.78.

ESI-MC Calculate. for C ₅₉ H ₈₃ N ₅ O ₁₅ : 1101.59. Found 1102.7 (M + H) ⁺ .

Example 36

Synthesis of Boc-Thr (Z-N (Me) -O (Me) -Tyr) -Ist-Val-Leu-Pro-OBn (SVPL7)

Following the procedure developed for the synthesis of SAPL7, starting from SVPLA1 (1.44 g, 2.37 mmol) and D1 (1.29 g, 2.37 mmol), the title compound (1.96 g, 75%) was obtained in the form of a white solid after purification by flash LC (silica gel, hex. gradient - EtOAc 2: 1 - 1: 3). Rf = 0.56 (EtOAc).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.83-0.95 (m, 15H), 1.0-1.22 (m, 4H), 1.23-1.44 (m, 9H) 1.60-1.65 (m, 1H), 1.87-2.01 (m, 4H), 2.09-2.20 (m, 3H), 2.77 (br s, 8H) 2.84-3.01 (m, 1H), 3.17-3.24 (m, 1H), 3.51-3.60 (m, 1H), 3.73 (s, 3H), 3 -80-3.90 (m, 2H), 4.03-4.15 (m, 1H), 4.25-4.40 (m, 2H), 4.40-4.52 (m, 1H) 4.70-4.80 (m, 2H), 5.00-5.26 (m, 4H), 5.34-5.36 (m, 1H), 5.58 (m, 1H), 6 75 (d, 2H, J = 7.8), 6.96-7.09 (m, 1H), 7.04 (d, 2H, J = 8.1), 7.04-7.12 ( m, 1H), 7.16-7.20 (m, 1H), 7.18-7.30 (m, 10H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.43, 13.63, 17.17, 18.35, 19.21, 21.40, 23.23, 24.49, 24.67, 26, 87, 28.10, 28.79, 30.48, 32.11, 33.84, 38.47, 40.37, 41.22, 46.80, 48.61, 55.04, 56.88, 57.95, 58.75, 59.25, 60.82, 66.87, 67.33, 69.40, 70.50, 76.44, 80.45, 113.58, 127.51, 127, 74, 127.88, 128.10, 128.25, 128.33, 128.46, 128.72, 129.64, 129.77, 135.35, 136.37, 156.77, 158.29, 169.83, 170.57, 171.3, 171.4, 172.81.

ESI-MC Calculate. for C ₅₉ H ₈₄ N ₆ O ₁₄ : 1100.60. Found (m / z): 1101.7 (M + H) ⁺ .

Example 37

Synthesis of Boc-Thr (N (Me) -O (Me) -Tyr) -Ist-Hip-Leu-Pro-OH (SAPL6)

Pd (OH) ₂ -C (20% Pd, 7.33 g, 40% m) is added to a solution of SAPL7 (18.33 g, 15.8 mmol) in THF (without stabilizer, 500 ml), degassed and purged with argon. / m). The mixture was stirred under an atmosphere of H ₂ (1 atm) for 20 hours, then filtered through a 0.45 μm Teflon filter and concentrated under reduced pressure to give a white solid. Toluene (30 ml) was added and concentrated again under reduced pressure and deep vacuum to give SAPL6 (14.78 g, quant.) As a white solid.

¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.79-1.08 (m, 18H), 1.09-1.18 (m, 3H), 1.26 (br s, 3H), 1 29 (d, J = 7.1, 3H), 1.47 (s, 9H), 1.50-1.66 (m, 3H), 1.84-1.94 (m, 1H), 1 90-2.28 (m, 4H), 2.35-2.50 (m, 4H), 2.30-2.35 (m, 1H), 2.44-3.18 (m, 4H) 2.60 (m, 3H), 3.53-3.61 (m, 1H), 3.77 (s, 3H), 3.88-4.07 (m, 4H), 4.12-4 72 (m, 4H), 5.18-5.24 (m, 1H), 5.24 (br s, 1H), 6.84 (d, J = 7.9, 2H), 7.08 (d, J = 8.0, 2H), 7.13 (d, J = 8.2, 1H), 7.18 (d, J = 8.2, 1H), 7.62-7.68 ( br.s., 1H).

m / z (FAB) 972.7 [(M + K) ⁺ , 33], 934.9 (M) ⁺ , 100].

Example 38

Synthesis of Boc-Thr (N (Me) -O (Me) -Tyr) -Ist-Aip-Leu-Pro-OH (SNPL6)

To a solution SNPL7 (130 mg, 0.11 mmol) in IPA: H ₂ O (2: 1, 4 ml: 2 ml) degassed and purged with argon, was added Pd (OH) ₂ -C (20% Pd, 45 mg, 35% m / m). The mixture was stirred under an atmosphere of H ₂ (1 atm) for 20 hours, then filtered through a 0.45 μm Teflon filter and concentrated under reduced pressure to give a white solid. IPA (10 ml) was added and concentrated again under reduced pressure and deep vacuum to give SNPL6 (100 mg, quant.) As a white solid.

ESI-MC Calculate. for C ₄₇ H ₇₆ N ₆ O ₁₃ : 932.55. Found 934.0 (M + H) ⁺ .

Example 39

Synthesis of Boc-Thr (N (Me) -O (Me) -Tyr) -Ist-Hiv-Leu-Pro-OH (SHPL6)

Follow the procedure developed for the synthesis of SAPL6 based on SHPL7 (1.045 g, 0.95 mmol). The title compound (825 mg, 99%) was obtained as a white solid.

ESI-MC Calcd for C ₄₄ H ₇₁ N ₅ O ₁₃ : 877.50. Found 878.5 (M + H) ⁺ .

Example 40

Synthesis of Boc-Thr (N (Me) -O (Me) -Tyr) -Ist-Val-Leu-Pro-OH (SVPL6)

Follow the procedure developed for the synthesis of SAPL6 based on SVPL7 (250 mg, 0.23 mmol). The title compound (195 mg, 97%) was obtained as a white solid.

ESI-MC Calculate. for C ₄₄ H ₇₂ N ₆ O ₁₂ : 876.56. Found (m / z): 877.5 (M + H) ⁺ .

Example 41

Synthesis of Cyclo-N (Me) -O (Me) -Tyr-O- (Boc-Thr) -Ist-Hip-Leu-Pro (SAPL5)

In a cooled (-5 ° C) 5 L reactor equipped with a mechanical stirrer containing ACN (3.2 L), HATU (14.436 g, 37.9 mmol) and HOAt (5.254 g, 38.6) were added under Ar atmosphere with stirring. mmol). Add SAPL6 (14.77 g, 15.8 mmol) dissolved in ACN (500 ml). NMM (3.65 ml, 33.18 mmol) was added dropwise using a syringe, keeping the temperature below -5 ° C. The resulting reaction mixture was allowed to warm to room temperature and stirred for 20 hours. The solvent was evaporated under reduced pressure. The crude product was distilled with EtOAc (500 ml) and the solution was filtered to remove a precipitate. The solution is washed successively with water. KHSO ₄ (2 × 500 ml), brine (500 ml), aq. NaHCO ₃ (500 ml, sat.) And rinsed with brine (500 ml). The organic phase was dried (Na ₂ SO ₄ ) and concentrated under reduced pressure to give a yellow solid (17.52 g), which was purified by flash chromatography (LC) (silica gel, hex.: EtOAc 3: 1 - 1: 1 ), obtaining SAPL5 (9.83 g, 68%) as a white solid. Rf = 0.60 (hex.: EtOAc 1: 3). [α] _D -209.4 (c 0.3, CHCl ₃ ).

IR (film, DCM) ν 3343, 2961, 2927, 2893, 1734, 1640, 1514, 1454, 1368, 1302, 1248, 1167, 1018 cm ^-1 .

¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.78 (d, J = 7.1, 3H), 0.85 (d, J = 7.0, 3H), 0.87 (d, J = 7.0, 3H), 0.89-0.93 (m, 9H), 1.10-1.20 (m, 1H), 1.20 (d, J = 6.4, 3H), 1, 30 (d, J = 6.9, 3H), 1.36 (m, 2H), 1.40 (m, 2H), 1.44 (s, 9H), 1.48-1.72 (m, 2H), 1.72-1.78 (m, 1H), 1.83-1.88 (m, 1H), 2.01-2.17 (m, 3H), 2.27-2.29 ( m, 1H), 2.47-2.53 (m, 1H), 2.53 (s, 3H), 2.93 (br.s, 1H), 3.14-3.19 (m, 2H) 3.34-3.37 (dd, J ₁ = 14.8, J2 = 4, l, 1H); 3.54-3.56 (dd, J ₁ = 10.5, J ₂ = 4, l , 1H), 3.58-3.63 (m, 1H), 3.68-3.72 (m, 1H), 3.78 (s, 3H), 3.94-3.98 (m, 1H ), 3.98 (q, J = 7.5, 1H), 4.07-4.11 (3d, J = 3.8, 1H), 4.57-4.61 (m, 2H), 4 77-4.81 (m, 1H), 4.97-4.98 (q, J = 3.5, 1H), 5.02 (d, J = 10.5, 1H), 5.18 ( d, J = 4.2, 1H), 6.81 (d, J = 8.5, 2H), 7.05 (d, J = 8.5, 2H), 7.19 (d, J = 10 2, 1H); 7.64 (d, J = 10.1, 1H).

¹³ C-NMR (75 MHz, CDCl ₃₎ δ 11.56, 14.68, 14.97, 15.27, 16.61, 18.45, 20.64, 23.50, 24.71, 24, 78, 26.92, 27.73, 27.94, 31.55, 33.94, 33.94, 38.27, 38.52, 40.64, 46.86, 49.54, 49.65, 55.16, 55.19, 55.84, 57.12, 65.96, 67.30, 71.00, 80.27, 81.41, 114.02, 130.22, 158.53, 168, 30, 169.31, 170.12, 170.29, 171.20, 172.38, 204.51.

m / z (FAB) 938.9 [(M + Na) ⁺ , 55], 916.9 [(M + H) ⁺ , 100]; m / z (FABHRMS) 916.532 120, C ₄₇ H ₇₃ N ₅ O ₁₃ requires (M + Na) ⁺ = 916.528 300.

Example 42

Synthesis of Cyclo-N (Me) -O (Me) -Tyr-O- (Boc-Thr) -Ist-Aip-Leu-Pro (SNPL5)

Following the procedure developed for the synthesis of SAPL5, starting from SNPL6 (100 mg, 0.11 mmol), the title compound (40 mg, 57%) was obtained as a white solid after flash LC (silica gel, EtOAc: hex. 4: 1 - pure EtOAc). Rf = 0.4 (EtOAc).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.76 (d, J = 6.8, 3H), 0.83-0.96 (m, 15H), 1.10-1.20 (m, 1H), 1.25 (d, J = 6.4, 3H), 1.27 (d, J = 6.3, 3H), 1.32 (d, J = 6.8, 3H), 1, 41 (m, 2H), 1.44 (s, 9H), 1.50-1.70 (m, 2H), 1.99-2.31 (m, 5H), 2.61 (s, 3H) 2.91-3.04 (m, 1H), 3.11-3.37 (m, 2H), 3.48-3.64 (m, 3H), 3.69-3.81 (m, 1H), 3.80 (s, 3H), 4.18 (m, 2H), 4.46-4.67 (m, 3H), 4.81 (t, J = 10.7, 1H), 5 01 (m, 1H), 6.85 (d, J = 8.3, 2H), 7.07 (d, J = 8.3.2H), 7.33 (d, J = 8.7, 1H), 7.65 (d, J = 9.2, 1H), 7.86 (d, J = 10.7, 1H).

ESI-MC Calculate. for C ₄₇ H ₇₄ N ₆ O ₁₂ 914.54. Found (m / z) 915.5 (M + H) ⁺ .

Example 43

Synthesis of Cyclo-N (Me) -O (Me) -Tyr-O- (Boc-Thr) -Ist-Hiv-Leu-Pro (SHPL5)

Following the procedure developed for the synthesis of SAPL5, starting from SHPL6 (2.45 g, 2.78 mmol), the title compound (2.1 g, 88%) was obtained as a white solid after crystallization of DCM / n-heptane (1 : 3). Rf = 0.33 (hex. EtOAc 1: 3).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.04 (m, 18H), 1.19 (m, 5H), 1.41 (s, 9H), 1.42 (m, 2H ), 1.63 (m, 1H), 1.77 (m, 1H), 1.90 (m, 1H), 2.00-2.22 (m, 3H), 2.44 (m, 1H) 2.58 (s, 3H), 2.95 (m, 1H), 3.14 (m, 1H), 3.26 (m, 1H), 3.36 (m, 1H), 3.58 ( m, 1H), 3.68 (m, 2H), 3.78 (s, 3H), 3.96 (m, 1H), 4.12 (m, 1H), 4.30 (m, 1H), 4.61 (m, 1H), 4.87 (m, 1H), 4.94 (m, 1H), 5.03 (m, 1H), 6.84 (d, J = 8.3, 2H) 7.07 (d, J = 8.3, 2H); 7.54 (d, J = 7.3, 1H); 7.69 (d, J = 6.4, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.38, 14.57, 15.15, 18.11, 18.45, 20.67, 23.40, 24.70, 26.91, 27, 89, 30.20, 33.57, 33.90, 38.51, 39.07, 46.62, 48.13, 55.16, 56.05, 56.23, 56.94, 65.67, 68.60, 71.13, 79.41, 80.01, 113.97, 129.69, 130.25, 155.75, 158.49, 168.51, 169.57, 170.24, 170, 94, 171.06, 173.59.

ESI-MC: Calculate. for C ₄₄ H ₆₉ N ₅ O ₁₂ 859.49. Found (m / z) 860.4 (M + H) ⁺ .

Example 44

Synthesis of Cyclo-N (Me) -O (Me) -Tyr-O- (Boc-Thr) -Ist-Val-Leu-Pro (SVPL5)

Following the procedure developed for the synthesis of SAPL5, starting from SVPL6 (90 mg, 0.1 mmol), 30 mg (35%) of the title compound was obtained as a white solid after purification by flash LC (silica, hex gradient. EtOAc from 1: 4 to 1:10). Rf = 0.35 (EtOAc).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.85-1.00 (m, 18H), 1.14-1.38 (m, 8H), 1.44 (s, 9H), 1.57 (m, 2H), 1.76-1.95 (m, 2H), 2.01-2.21 (m, 2H), 2.33 (dd, J ₁ = 7.3, J ₂ = 14, 7, 1H), 2.53 (m, 1H), 3.57 (s, 3H), 3.17 (dd, J ₁ = 10.7, J ₂ = 14.7, 1H), 3.35 (dd, J ₁ = 4.4, J ₂ = 14.2, 1H), 3.56 (dd, J ₁ = 3.9, J ₂ = 10.3, 1H), 3.59-3.77 (m, 4H), 3.78 (s, 3H), 4.06 (dt, J ₁ = 3.9, J ₂ = 9.3, 1H), 4.33 (dd, J ₁ = 2, 9, J ₂ = 9.3, 1H), 4.38 (dd, J ₁ = 6.8, J ₂ = 10.3, 1H), 4.58 (dd, J ₁ = 5.4, J ₂ = 7.3 , 1H), 4.79 (t, J = 10.3, 1H), 4.98 (d, J = 9.3, 1H), 5.03 (dd, J ₁ = 2.4, J ₂ = 6.3, 1H), 6.81 (br s, 1H), 6.84 (d, J = 8.3, 2H), 7.08 (d, J = 8.3, 2H), 7, 24 (d, J = 9.8, 1H); 7.54 (d, J = 9.8, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.48, 14.50, 15.20, 18.86, 19.44, 21.03, 23.64, 24.83, 24.96, 26, 97, 28.01, 28.135, 30.21, 33.53, 33.96, 38.56, 38.61, 41.05, 41.66, 46.84, 48.53, 55.24, 55, 51, 56.31, 57.14, 59.75, 65.85, 70.60, 80.52, 114.11, 129.75, 130.33, 156.77, 158.67, 168.53, 170.21, 170.29, 170.73, 171.02, 174.34.

ESI-MC Calculate. for C ₄₄ H ₇₀ N ₆ O ₁₁ : 858.51. Found (m / z): 859.5 (M + H) ⁺ .

Example 45

Synthesis of Cyclo-N (Me) -O (Me) -Tyr-O- (Thr) -Ist-Hip-Leu-Pro (SAPL4)

In a flask containing SAPL5 (8.79 g, 9.6 mmol) in bezv. dioxane (93 ml), add a solution of hydrochloric acid in bezv. dioxane (5.3 n, 122 ml, 647 mmol). The resulting solution was stirred at room temperature for 8 hours or until the starting material completely disappeared. After completion of the reaction, the solution was concentrated under reduced pressure. The residue was diluted with CHCl ₃ (100 ml) and concentrated again. The foamy crude product was evaporated together with CHCl ₃ / hex., Which gives SAPL4 (8.17 g, 100% yield) as a white solid.

m / z (FAB) 838.3 [(M + Na) ⁺ , 28], 816.3 [(M + H) ⁺ , 100].

Example 46

Synthesis of cyclo-N (Me) -O (Me) -Tyr-O- (Thr) -Ist-Aip-Leu-Pro (SNPL4:

Following the procedure developed for the synthesis of SAPL4, starting from SNPL5 (40 mg, 43 μmol), the title compound (36 mg, quant.) Was obtained as a white solid after precipitation with Et ₂ O.

ESI-MC: Calculate. for C ₄₂ H ₆₅ N ₅ O ₁₁ 815.47. Found m / z 825.5 (M) ⁺ .

Example 47

Synthesis of cyclo-N (Me) -O (Me) -Tyr-O- (Thr) -Ist-Hiv-Leu-Pro (SNPL4)

Follow the procedure developed for the synthesis of SAPL4 based on SHPL5 (500 mg, 0.58 mmol). The title compound (440 mg, quant.) Was obtained as a white solid after precipitation of Et ₂ O.

ESI-MC: Calculate. for C ₃₉ H ₆₁ N ₅ O ₁₀ 759.44. Found m / z 760.4 (M + H) ⁺ .

Example 48

Synthesis of Cyclo-N (Me) -O (Me) -Tyr-O- (Thr) -Ist-Val-Leu-Pro (SVPL4)

Follow the procedure developed for the synthesis of SAPL4 based on SVPL5 (25 mg, 29 μmol). The title compound (22 mg, quant.) Was obtained as a white solid after coevaporation with MTBE.

ESI-MC Calculate. for C ₃₉ H ₆₂ N ₆ O ₉ : 758.5. Found (m / z): 759.5 [(M + H)] ⁺ .

Example 49

Z-N-Methyl-D-Leucine (H1)

Ref: Coggins, J.R .; Benoiton, N.L. Can. J. Chem 1971, 49, 1968.

To a stirred solution of ZD-Leu-OH (10.32 g, 38.9 mmol) in bezv. THF (120 ml) at 0 ° C. under an Ar atmosphere is added dropwise through a syringe of iodomethane (8.55 ml, 136.1 mmol). Then sodium hydride (4.80 g, 120.6 mmol, 60% dispersion in mineral oil) was added portionwise while maintaining the temperature at 0 ° C. The reaction mixture was stirred at room temperature for 24 hours. Traces of the solvent were removed under reduced pressure and the residue dissolved in EtOAc (120 ml) and extracted with aq. NaHCO ₃ (300 ml, sat.). The aqueous phase is washed with EtOAc (2 × 100 ml). The aqueous phase is cooled, solid citric acid is added to pH 1-2 and the solution is extracted with EtOAc ( 4 × 250 ml), dried (Na ₂ SO ₄ ), filtered and concentrated.The product is crystallized in EtOAc-heptane (1: 3) to give H1 (7.84 g, 7 2%) as a white crystalline solid, mp: 71-72 ° C. [Α] _D ²⁵ +23 (s 1, EtOH).

Example 50

Synthesis of Z-Didemnin A (SAPL3)

Into a flask containing HATU (8.76 g, 23.0 mmol), HOAt (3.17 g, 23.1 mmol) SAPL4 (7.09 g, 15.8 mmol) and H1 (3.486 g, 12.5 mmol), add bezv. DCM (100 ml) and anv. DMF (50 ml) in an Ar atmosphere and the solution was stirred at -5 ° C (ice bath). NMM (2.3 ml, 21.0 mmol) was added dropwise via syringe while maintaining the temperature at -5 ° C. The resulting reaction mixture was stirred at -5 ° C for 2 hours, then allowed to warm to room temperature for another 14 hours. The solvent was evaporated under reduced pressure. The crude product was distilled with EtOAc (100 ml) and the solution was filtered to remove some precipitate. The solution was washed successively with aq. KHSO ₄ (2 × 100 ml, 10%), brine (100 ml), aq. NaHCO ₃ (100 ml, sat.) And rinsed with brine (100 ml). The organic phase was dried (Na ₂ SO ₄ ) and concentrated under reduced pressure to obtain a yellow solid, which was purified by flash LC (silica gel, hex. Gradient: EtOAc 2: 1 - 1: 1) to give SAPL3 (7.98 g, 89% yield) as a white solid. Rf = 0.18 (hex. / EtOAc 1: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.79-1.00 (m, 24H), 1.10-2.25 (m, 10H), 1.18 (d, J = 6.3, 3H), 1.25 (s, 3H), 1.32 (d, J = 6.8, 3H), 2.28-2.34 (m, 1H), 2.49 (dd, J ₁ = 10 7, J ₂ = 17.0, 1H), 2.54 (s, 3H), 2.83 (s, 3H), 2.95 (m, 1H), 3.02-3.24 (m, 2H), 3.31-3.40 (dd, J ₁ = 3.9, J ₂ = 14.1, 1H), 3.53-3.64 (m, 2H), 3.65-3.75 (m, 1H), 3.78 (s, 3H), 3.92-4.20 (m, 3H), 4.58 (t, J = 4.8, 1H), 4.75-4.85 (m, 3H), 5.00 (m, 1H), 5.12-5.26 (m, 3H), 6.84 (d, J = 8.3, 2H), 6.86 (br s , 1H), 7.07 (d, J = 8.3, 2H), 7.21-7.44 (m, 6H), 7.92 (d, J = 8.3, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.57, 14.05, 15.18, 16.73, 18.52, 20.83, 22.62, 22.96, 23.66, 24, 55, 24.81, 25.03, 25.28, 26.90, 27.86, 28.95, 31.28, 31.81, 33.91, 34.02, 38.51, 38.61, 47.03, 49.61, 55.21, 55.38, 55.50, 57.28, 66.10, 67.65, 67.93, 70.47, 81.44, 114.09, 127, 83, 128.46, 129.74, 130.29, 158.60, 168.36, 169.59, 170.28, 171.20, 172.22, 204.80.

ESI-MC Calculate. for C ₅₇ H ₈₄ N ₆ O ₁₄ 1076.60. Found m / z 1077.6 (M + H) ⁺ .

Example 51

Synthesis of [Aip] ³ -Z-Didemnin A (SNPL3)

Following the procedure developed for the synthesis of SAPL3, starting from SNPL4 (35 mg, 41 μmol) and H1 (17 mg, 61 μmol), the title compound (36 mg, 81%) was obtained as a white solid after flash chromatography ( silica gel, hex gradient - EtOAc from 1: 4 to pure EtOAc). Rf = 0.30 (EtOAc).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.74 (d, J = 6.3, 3H), 0.80-1.00 (m, 21H), 1.10-2.25 (m, 10H), 1.21 (d, J = 5.8, 3H), 1.24 (s, 3H), 1.34 (d, J = 6.3, 3H), 2.61 (s, 3H) , 2.86 (s, 3H), 3.12-3.25 (dd, J ₁ = 11.2, J ₂ = 14.1, 1H), 3.27-3.36 (dd, J ₁ = 4.3, J ₂ = 14.1, 1H), 3.52-3.63 (m, 3H), 3.69-3.81 (m, 2H), 3.80 (s, 3H), 4 09 (m, 3H), 4.47-4.63 (m, 3H), 4.76-4.92 (m, 2H), 5.00 (m, 1H), 5.08 (m, 1H) ), 5.18 (s, 2H), 6.85 (d, J = 8.3, 2H), 6.97 (d, J = 6.97, 1H), 7.07 (d, J = 8 , 3, 2H), 7.35 (br s, 5H), 7.48 (d, J = 8.3, 1H), 7.67 (d, J = 8.3, 1H), 7.87 (d, J = 10.2, 1H).

ESI-MC Calculate. for C ₅₇ H ₈₅ N ₇ O ₁₃ : 1075.62. Found m / z: 1076.6 (M + H) ⁺ .

Example 52

Synthesis of [Hiv] ³ -Z-Didemnin A (SHPL3)

Following the procedure developed for the synthesis of SAPL3, starting from SHPL4 (116 mg, 0.15 mmol) and H1 (63 mg, 0.23 mmol), the title compound (86 mg, 52%) was obtained as a white solid after flash LC purification (silica gel, hex gradient. EtOAc from 1: 1 to 1: 2). Rf = 0.27 (hex. - EtOAc 1: 2).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.80-1.08 (m, 24H), 1.18 (m, 3H), 1.21 (m, 4H), 1.58 (m, 2H ), 1.74 (m, 1H), 1.80-2.42 (m, 6H), 2.56 (s, 3H), 2.80 (s, 3H), 2.88 (m, 1H) , 3.15 (m, 1H), 3.32 (m, 1H) 3.60 (m, 3H), 3.78 (s, 3H), 3.83 (m, 1H), 3.98 (m , 1H), 4.42 (m, 1H), 4.58 (m, 1H), 4.75 (m, 1H), 4.84 (m, 1H), 4.92 (d, J = 3, 8, 1H), 5.00 (m, 1H), 5.20 (m, 2H), 6.65 (d, J = 6.3, 1H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8.3, 2H), 7.34 (m, 5H), 7.50 (d, J = 6.7, 1H), 7.75 (d, J = 7.2, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.58, 14.21, 15.39, 17.64, 18.63, 20.74, 21.88, 22.84, 23.48, 24, 22, 24.75, 27.05, 27.84, 29.34, 29.99, 33.42, 33.86, 35.81, 38.52, 39.37, 46.63, 48.14, 55.13, 55.47, 55.53, 55.87, 56.92, 65.68, 67.72, 68.60, 70.61, 79.09, 113.95, 127.71, 127, 86, 128.35, 129.63, 130.22, 158.48, 168.46, 169.36, 169.84, 170.29, 170.93, 171.00, 173.73.

ESI-MC Calculate. for C ₅₄ H ₈₀ N ₆ O ₁₃ 1020.58. Found m / z 1021.5 (M + H) ⁺ .

Example 53

Synthesis of [Val] ³ -Z-Didemnin A (SVPL3)

Following the procedure developed for the synthesis of SAPL3, starting from SVPL4 (20 mg, 25 μmol) and SAPLH1 (11 mg, 37.5 μmol), the title compound (19 mg, 72%) was obtained as a white solid after flash purification LC (silica gel, hex. Gradient - EtOAc from 1: 1 to 1: 5). Rf = 0.44 (EtOAc).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.85-1.06 (m, 21H), 1.10-1.4 (m, 5H), 1.16 (d, J = 6.6, 3H), 1.50-1.63 (m, 6H), 1.72-1.87 (m, 2H), 1.88-2.40 (m, 6H), 2.58 (s, 3H) 2.86 (s, 3H), 3.17 (dd, J ₁ = 10.5, J ₂ = 14.2, 1H), 3.36 (dd, J ₁ = 3.9, J2 = 14, 2, 1H), 3.43 (br s, 1H), 3.51-3.72 (m, 4H), 3.79 (s, 3H), 3.98-4.16 (m, 1H) 4.40-4.47 (m, 2H); 4.58 (dd, J ₁ = 5.7; J2 = 7.8; 1H); 4.67-4.85 (m, 2H); 4 80-5.09 (m, 1H), 5.16 (d, J = 12.4, 1H), 5.24 (d, J = 12.4, 1H), 6.84 (d, J = 8.4, 1H), 6.90-6.94 (br s, 1H), 7.09 (d, J = 8.4, 1H), 7.28-7.50 (m, 6H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.99, 14.37, 15.70, 18.55, 19.82 21.40, 21.99, 22.81, 23.17, 23.98 , 24.62, 25.00, 25.24, 27.28, 28.26, 29.93, 30.23, 33.58, 34.10, 36.16, 38.78, 41.98, 47 , 10, 48.79, 54.73, 55.45, 56.62, 57.40, 59.51, 66.02, 68.18, 70.37, 71.03, 114.30, 128.00 , 128.27, 128.72, 129.86, 130.56, 136.49, 158.28, 158.85, 168.75, 169.27, 170.40, 170.75, 171.04, 173 91, 175.03.

ESI-MC Calculate. for C ₅₄ H ₈₁ N ₇ O ₁₂ : 1019.59. Found (m / z): 1020.5 (M + H) ⁺ .

Example 54

Synthesis of Didemnin A (SAPL2)

To a solution of SAPL3 (6.59 g, 6.1 mmol) in THF (without stabilizer, 262 ml) degassed and purged with argon was added Pd (OH) ₂ -C (20%, 3.29 g, 50% m / m). The mixture was stirred under an atmosphere of H ₂ (1 atm) for 20 hours, then filtered through a 0.45 μm Teflon filter and concentrated under reduced pressure to give a white solid. CHCl ₃ (2 × 25 ml) was added and the mixture was again concentrated under reduced pressure to give SAPL2 (5.51 g, 96%) as a white solid. Rf = 0.22 (CHCl ₃ -t-BuOH 90:10).

¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.82-0.92 (m, 24H), 1.11-1.19 (m, 1H), 1.22 (d, J = 6.9, 3H), 1.32 (d, J = 6.8, 3H), 1.30-1.35 (m, 1H), 1.35-1.63 (m, 6H), 1.71-1, 81 (m, 2H), 1.93-2.07 (m, 1H), 2.07-2.18 (m, 2H), 2.28-2.34 (m, 1H), 2.49- 2.52 (dd, J ₁ = 11, J ₂ = 10.5, 1H), 2.54 (s, 3H), 2.72 (br s, 3H), 2.79 (br s, 3H ), 2.86-2.94 (br s, 1H), 2.72-2.79 (br d, J = 10.5, 1H), 3.15-3.18 (dd, J ₁ = 14.5, J ₂ = 10.5, 1H), 3.33-3.36 (dd, J ₁ = 14.5, J ₂ = 4.5, 1H), 3.54-3.57 ( dd, J ₁ = 10.5, J ₂ = 4.5, 1H), 3.56-3.61 (m, 1H), 3.78 (s, 3H), 3.96-4.00 (m , 1H), 4.03-4.08 (m, 1H), 4.11-4.80 (br.s, 1H), 4.56-4.62 (m, 1H), 4.68-4 81 (m, 3H), 4.99-5.01 (q, J = 3.5, 1H), 5.16 (br s, 1H), 6.83 (d, J = 8.5, 2H), 6.95 (br.s, 1H), 7.07 (d, J = 8.5, 2H), 7.21-7.25 (br.s, 1H), 7.95 (br. s, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.55, 14.95, 15.26, 16.82, 18.56, 20.89, 22.00, 23.08, 23.76, 24, 58, 24.85, 25.10, 27.12, 29.35, 29.35, 29.65, 29.69, 31.36, 33.96, 34.14, 38.51, 38.64, 40.14, 55.38, 55.56, 57.31, 66.17, 67.85, 70.58, 80.96, 80.98, 81.57, 114.12, 130.33, 158, 63, 168.41, 169.33, 169.70, 170.38, 171.24, 172.28, 172.28, 172.93, 204.83.

m / z (FAB) 944.2 [(M + H) ⁺ , 100].

Example 55

Synthesis of [Aip] ^3- didemnin A (SNPL2)

To a solution of SNPL3 (33 mg, 35 μmol) in an IPA / H ₂ O mixture (2 ml: 1 ml), degassed and purged with argon, Pd (OH) ₂ -C (20%, 20 mg, 60% m / m) was added ) The mixture was stirred under an atmosphere of H ₂ (1 atm) for 20 hours, then filtered through a 0.45 μm Teflon filter and concentrated under reduced pressure to give a white solid. IPA (2 × 5 ml) was added and the solution was again concentrated under reduced pressure to obtain SNPL2 (32 mg, 97%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.78 (d, J = 6.8, 3H), 0.82-0.92 (m, 21H), 1.11-2.58 (m, 21H), 2.41 (s, 3H), 2.62 (s, 3H), 2.75-3.00 (m, 4H), 3.15-3.18 (dd, J ₁ = 10.7 , J ₂ = 14.7, 1H), 3.33-3.36 (dd, J ₁ = 14.5, J ₂ = 4.2, 1H), 3.51-3.78 (m, 3H) 3.78 (s, 3H), 3.92 (m, 1H), 4.01-4.20 (m, 2H), 4.50 (t, J = 4.8, 1H), 4.59 (t, J = 6.3, 1H), 4.75-4.91 (m, 2H), 5.05 (m, 1H), 6.84 (d, J = 8.3, 2H), 7 07 (d, J = 8.3, 2H), 7.70 (d, J = 5.8, 1H), 7.78 (d, J = 9.7, 1H), 7.89 (d, J = 6.3, 1H); 8.14 (d, J = 7.8, 1H).

ESI-MC Calculate. for C ₄₉ H ₇₉ N ₇ O ₁₁ 941.58. Found m / z 942.7 (M + H) ⁺ .

Example 56

Synthesis of [Hiv] ^3- didemnin A (SHPL2)

Following the procedure developed for the synthesis of SAPL2, starting from SHPL3 (86 mg, 0.08 mmol), the title compound (73 mg, 97%) was obtained as a white solid. Rf = 0.36 (CHCl ₃ / MeOH 95: 5).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.02 (m, 24H), 1.12-2.42 (m, 16H), 2.54 (s, 3H), 2.64 (s, 3H), 2.95 (m, 1H), 3.15 (m, 1H), 3.35 (m, 1H), 3.52-3.90 (m, 5H), 3.78 ( s, 3H), 4.04 (m, 1H), 4.38 (m, 1H), 4.48 (m, 1H), 4.57 (m, 1H), 4.88 (m, 1H), 4.91 (d, J = 5.3, 1H), 5.22 (m, 1H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8.3 , 2H), 7.54 (d, J = 9.2, 1H), 7.60 (d, J = 9.4, 1H), 8.68 (d, J = 6.2, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.78, 14.07, 16.13, 17.90, 18.56, 20.98, 21.74, 22.94, 23.60, 24, 44, 24.78, 25.06, 27.17, 27.92, 30.09, 33.34, 33.89, 38.71, 40.30, 46.86, 48.22, 54.99, 55.23, 56.97, 57.21, 65.81, 68.53, 70.37, 79.47, 114.03, 129.76, 130.29, 158.57, 168.18, 169, 40, 169.86, 170.20, 170.92, 174.16.

ESI-MC Calculate. for C ₄₆ H ₇₄ N ₆ O ₁₁ 886.54. Found m / z 887.2 (M + H) ⁺ .

Example 57

Synthesis of [Val] ^3- didemnin A (SVPL2)

Following the procedure developed for the synthesis of SAPL2, starting from SVPL3 (19 mg, 18 μmol), the title compound (16 mg, 97%) was obtained as a white solid. Rf = 0.36 (CHCl ₃ / MeOH 95: 5).

¹ H-NMR (300 MHz, CDCl ₃ ) δ -0.82-1.02 (m, 21H), 1.85-1.32 (m, 9H), 1.42-1.83 (m, 7H ), 1.87-2.23 (m, 8H), 2.34 (br.s, 3H), 2.60 (br.s, 1H), 2.85 (dd, J ₁ = 5.7, J ₂ = 7.8, 1H), 3.16 (dd, J ₁ = 10.8, J ₂ = 14.2, 1H), 3.37 (dd, J ₁ = 4.2, J ₂ = 14 , 2, 1H), 3.48 (br.s, 1H), 3.57-3.68 (m, 4H), 3.79 (s, 3H), 3.95-4.15 (m, 2H) ), 4.45-4.52 (m, 2H), 4.61 (dd, J ₁ = 4.8, J ₂ = 7.8, 1H), 4.77 (t, J = 9, 9, 1H), 5.02-5.15 (m, 1H), 6.84 (d, J = 8.4, 1H), 7.09 (d, J = 8.4, 1H), 7.59- 7.62 (m, 2H); 7.78-7.81 (m, 2H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.98, 14.46, 16.14, 18.50, 19.83, 21.50, 22.55, 22.81, 22.95, 23, 46, 23.98, 27.33, 28.27, 28.33, 29.94, 33.71, 34.16, 38.94, 41.93, 42.11, 45.72, 47.18, 48.94, 53.64, 55.03, 55.50, 56.72, 57.55, 59.22, 59.56, 66.16, 70.71, 114.36, 118.53, 120, 85, 130.02, 130.58, 168.71, 169.74, 170.23, 171.18, 175.10.

ESI-MC Calculate. for C ₄₆ H ₇₅ N ₇ O ₁₀ : 885.56. Found (m / z): 856.5 (M + H) ⁺ .

Example 58

Synthesis of Pyr-Pro-OBn (F2)

To a solution of H-Pro-OBn-HCl (10.0 g, 41.3 mmol) in anv. DMF (50 ml) at 0 ° C in an Ar atmosphere, NMM (4.55 ml, 43.8 mmol) was added dropwise through a syringe, maintaining the temperature at 0 ° C. Then, HOBt (18.98 g, 124 mmol) was added in portions. After 15 minutes, pyruvic acid (8.61 g, 97.79 mmol) dissolved in anvil. DCM (10 ml) is added dropwise through a syringe, keeping the temperature below 3 ° C. Finally, DCC (22.17 g, 107.46 mmol) dissolved in DCM (80 ml) was added dropwise using a compensation funnel. The mixture was allowed to warm to room temperature (2 hours) and stirred for another 12 hours. The reaction mixture was filtered to remove a precipitate and the solution was concentrated under reduced pressure. The residue was dissolved in EtOAc (500 ml) and washed successively with aq. KHSO ₄ (100 ml, 10%), aq. NaHCO ₃ (400 ml, sat.), Brine (400 ml), dried (Na ₂ SO ₄ ) and filtered. The residual solvent was distilled off under reduced pressure, and the residue was distilled with ACN (100 ml), cooled to -30 ° C for 2 hours and filtered to remove excess N, N-dicyclohexylcarbamide. The resulting brown oil (15.82 g) was purified by flash LC (silica gel, deg. Hex. - hex.: EtOAc 2: 1) to give F2 (9.06 g, 66% yield) as a white solid. Rf = 0.25 (hex.: EtOAc 2: 1).

IR (film, DCM) ν 3035, 2956, 2884, 1744, 1717, 1645, 1499, 1443, 1383, 1352, 1273, 1175, 1092 cm ^-1 .

¹ H-NMR (200 MHz, CDCl ₃ ) δ 1.75-2.40 (m, 4H), 2.37 (s, 3H), 2.44 (s, 3H), 3.45-3.82 (m, 2H), 4.52-4.61 (m, 1H), 4.88-4.97 (m, 1H), 5.14-5.15 (m, 2H), 5.17-5 20 (m, 2H); 7.34 (br s, 5H).

¹³ C-NMR (50 MHz, CDCl ₃ ) δ 22.11, 25.22, 26.5, 27.10, 28.53, 31.48, 47.53, 44.81, 59.76, 67, 02, 67.31, 128.11, 128.64, 135.24, 170.1, 170.2, 198.0.

m / z (CI) 293 [(M + NH ₄ ) ⁺ , 100]. Anal calcd. C ₁₅ H ₁₇ NO ₄ : C, 65.44; H, 6.22; N, 5.08. Found: C, 65.04; H, 6.01; N, 5.11.

Example 59

Synthesis of Pyr-Pro-OH (F1)

A solution of F2 (8.63 g, 31.34 mmol) and palladium on activated carbon (10%, 86 mg, 10% m / m) in degassed MeOH (125 ml) were placed in a Parr reactor to operate at high pressure and purged with gaseous nitrogen (2 × 30 psi). The reaction mixture was isolated in a hydrogen atmosphere (30 psi) and stirred at 23 ° C. for 4.5 hours. The mixture was filtered through a 0.45 μm Teflon filter and concentrated under reduced pressure. The residue (6.29 g) was applied to SiO ₂ and the column was filled on top (LC 5.5 × 10.0 cm) and hex.: EtOAc 1: 2 was eluted to give F1 (4.64 g, 80%) as white solid matter. Rf = 0.22 (hex.: EtOAc 1: 1). [α] _D ²⁰ -92.4 (c 0.12, CHCl ₃ ). Mp 67-69 ° C.

IR (film, DCM) ν 3450-3000, 2961-2870, 1719, 1643, 1615, 1452, 1354, 1205, 1175, 1094, 1018 cm ^-1 .

¹ H-NMR (200 MHz, CDCl ₃ ) δ 1.85-2.45 (m, 4H), 2.43 and 2.47 (s, 3H), 3.42-3.85 (m, 2H) 4.52-4.61 (m, 1H); 4.88-4.97 (m, 1H); 7.21-7.40 (br s, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 22.03, 25.23, 26.48, 27.00, 28.23, 31.44, 47.57, 48.37, 59.61, 59, 39, 162.47, 162.52, 175.04, 176.29, 197.18.

m / z (CI) 220 [(M + N ₂ H ₇ ) ⁺ , 15], 203 [(M + NH ₄ ) ⁺ , 100], 186 [(M + H) ⁺ , 16]. Anal calculated C ₈ H ₁₁ NO ₄ : C, 51.88; H, 5.99; N, 7.56. Found: C, 52.13; H, 5.85; N, 7.65.

Example 60

Aplidine Synthesis (SAPL1)

To a cooled (3 ° C) solution of F1 (4.908 g, 26.5 mmol) in anv. DCM (40 ml) was added, under nitrogen, a solution of DIPCDI (1.806 mg, 14.3 mmol) in DCM (10 ml) and the solution was stirred at 3 ° C for 60 minutes Then a solution of SAPL2 (5.0 g, 5.3 mmol) in DCM (50 ml) was transferred via cannula into the first solution under nitrogen pressure. After 90 hours (4 days), aq. HCl (50 ml, 0.1 N) and the reaction mixture was stirred for 15 minutes. Then the organic layer was decanted and partitioned between KHSO ₄ (50 ml, 5%), aq. NaHCO ₃ (50 ml, 5%) and brine (25 ml). The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The resulting light yellow solid was purified by flash LC (Lichroprep RP-18, 40-63 μm, MeOH: H ₂ O: TFA gradient from 70: 30: 0.1 to 90: 10: 0.1), giving SAPL1 ( 5.4 g, 93%) (mixture of rotamers). Rf = 0.40 and 0.28 (DCM: AcOEt, 2: 3), 0.52 and 0.45 (CHCl ₃ -MeOH, 9.5: 0.5); [α] _D -95.9 (c 1.8, CHCl ₃ ).

¹ H-NMR (500 MHz, CDCl ₃ ) δ 0.84-0.93 (m, 24H), 1.16-1.70 (m, 9H), 1.72-1.81 (m, 1H) 1.81-1.90 (m, 1H), 1.90-2.24 (m, 6H), 2.30-2.39 (m, 1H), 2.49 (s, 3H) 2, 51 (s, 3H), 2.55 (s, 3H), 2.52-2.64 (m, 1H), 2.85 (br.s, 1H), 2.94 (br.s, 1H) , 3.09 (s, 3H), 3.13 (s, 3H), 3.15-3.18 (m, 1H), 3.21-3.26 (dd, J ₁ = 15.8, J ₂ = 6.1, 1H), 3.32-3.36 (dd, J ₁ = 14.5, J ₂ = 4.1, 1H), 3.54-3.60 (m, 1H), 3 66-3.72 (m, 1H), 3.78 (s, 3H), 3.80-3.87 (m, 1H), 3.96-3.99 (m, 1H), 4.03 -4.11 (m, 2H), 4.15-4.23 (2q, J = 7.5, 1H), 4.55-4.57 (2d, J ₁ = 5.5, J ₂ = 2 , 2, 1H), 4.59-4.62 (t, 1H), 4.56-4.64 (dd, J ₁ = 6.5, J ₂ = 2.5, 1H), 4.68- 4.71 (t, 1H), 4.76-4.81 (t, 1H), 5.10-5.18 (m, 1H), 5.17 (d, J = 3.5, 1H), 5.18 (d, J = 3.5, 1H), 5.27-5.31 (m, 2H), 6.82 (d, J = 8.5, 2H), 6.83 (d, J = 8.5, 2H), 7.05 (d, J = 8.5, 2H), 7.06 (d, J = 8.5, 2H), 7.02 (d, J = 7.1, 1H), 7.16 (d, J = 9.5, 1H), 7.17 (d, J = 9.5, 1H), 7.59 (d, J = 5.5, 1H), 7, 77 (d, J = 9.5, 1H), 7 83 (d, J = 9.4, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.63, 11.68, 14.11, 14.70, 15.26, 15.30, 16.00, 16.20, 16.88, 16, 93, 18.62, 18.85, 20.89, 20.94, 21.62, 21.36, 23.44, 23.57, 23.84, 23.93, 24.66, 24.77, 24.85, 25.02, 26.22, 26.34, 27.09, 27.6, 27.06, 27.30, 27.95, 27.99, 29.33, 29.69, 31, 31, 31.37, 33.97, 34.06, 36.02, 36.45, 38.68, 38.76, 41.01, 41.15, 47.00, 48.42, 48.48, 48.86, 49.20, 49.51, 54.65, 54.75, 55.26, 55.58, 55.61, 57.14, 57.27, 57.47, 57.79, 66, 24, 67.80, 67.99, 70.34, 70.67, 81.0, 81.52, 114.10, 130.31, 156.0, 158.65, 161.1, 161.60, 168.20, 169.53, 169.59, 170.45, 171.25, 171.80, 171.95, 172.26, 172.33, 197.5, 204.80, 204.85.

m / z (FAB) 1132.6 [(M + Na) ⁺ , 42], 1110.8 [(M + H) ⁺ , 100].

Example 61

Synthesis of [Aip] ³ -aplidine (SNPL1)

Follow the procedure developed for the synthesis of SAPL1 based on SNPL2 (10 mg, 10.6 μmol) and F1 (10 mg, 54 μmol). The title compound (8 mg, 68%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, isotrate ratio ACN / H ₂ O 85:15 (flow: 7 ml / min, 250 × 21 mm, 270 nm, t _R = 10.5 and 12.0 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.80-1.03 (m, 24H), 1.11-1.70 (m, 9H), 1.72-1.81 (m, 1H) 1.81-1.90 (m, 1H), 1.90-2.24 (m, 6H), 2.30-2.39 (m, 1H), 2.53 (s, 3H) 2, 55 (s, 3H), 2.65 (s, 3H), 2.52-2.66 (m, 2H), 2.94 (m, 1H), 3.07 (s, 3H), 3.11 (s, 3H), 3.15-3.18 (m, 1H), 3.22-3.31 (dd, J ₁ = 4.3, J ₂ = 15.1, 1H), 3.54- 3.60 (m, 2H), 3.67-3.92 (m, 2H), 3.80 (s, 3H), 3.98 (m, 1H), 4.13-4.29 (m, 3H), 4.45-4.75 (m, 4H), 4.81 (t, J = 9.7, 1H), 5.09 (m, 1H), 5.18 (m, 1H), 5 26-5.44 (m, 3H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8.3, 2H), 7.30 (d, J = 8.3, 1H), 7.36 (d, J = 8.3, 1H), 7.68 (d, J = 9.7, 1H), 7.87 (d, J = 4.3, 1H ), 8.09 (d, J = 9.7, 1H), 8.28 (d, J = 10.2, 1H).

ESI-MC Calculate. for C ₅₇ H ₈₈ N ₈ O ₁₄ : 1108.64. Found (m / z): 1110.3 (M + H) ⁺ .

Example 62

Synthesis of [Hiv] ³ -aplidine (SHPL1)

Follow the procedure developed for the synthesis of SAPL1, starting from SHPL2 (72 mg, 0.081 mmol) and F1 (75 mg, 0.405 mmol). The title compound (68 mg, 79%) was obtained as a white solid after purification by flash LC (Lichroprep RPC18, ACN / H ₂ O / TFA gradient from 70: 30: 0.5 to 90: 10: 0.5) . Rf = 0.49 (ACN / H ₂ O / TFA 90: 10: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.80-1.10 (m, 24H), 1.12-1.50 (m, 18H), 1.50-2.30 (m, 6H) 2.42 (m, 1H), 2.53 (s, 3H), 2.55 (s, 3H), 2.57 (s, 3H), 2.96-3.40 (m, 3H), 3.05 (s, 3H), 3.10 (s, 3H), 3.63 (m, 5H), 3.78 (s, 3H), 3.90 (m, 1H), 4.01 (m , 1H), 4.30 (m, 1H), 4.63 (m, 1H), 4.69 (m, 1H), 4.86 (m, 1H), 5.02 (d, J = 4, 8, 1H), 5.09 (m, 1H), 5.20 (m, 1H), 5.30 (m, 1H), 6.83 (d, J = 8.3, 2H), 6.89 (d, J = 6.3, 1H), 7.07 (d, J = 8.3, 2H), 7.29 (d, J = 9.7, 1H), 7.34 (m, 2H) 7.43 (d, J = 5.3, 1H); 7.74 (d, J = 9.7, 1H); 7.80 (d, J = 10.2, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 12.06, 14.22, 14.30, 16.49, 16.76, 17.84, 19.17, 21.08, 21.41, 21, 54, 22.54, 23.79, 23.94, 24.05, 24.16, 24.82, 24.96, 25.05, 25.98, 26.45, 27.37, 27.57, 28.21, 28.59, 30.31, 30.83, 31.56, 31.62, 33.74, 34.24, 36.02, 36.25, 38.91, 38.96, 39, 46, 39.86, 46.92, 48.44, 48.71, 49.06, 54.95, 55.49, 57.16, 57.68, 58.23, 59.13, 66.16 66.28, 69.10, 70.83, 71.14, 79.12, 114.31, 129.96, 130.12, 130.59, 158.86, 168.69, 168.81, 169, 75, 169.82, 170.18, 170.45, 170.52, 170.69, 170.84, 171.21, 171.28, 172.47, 173.17, 174.66, 174.82, 197.63, 201.38.

ESI-MC Calculate. for C ₅₄ H ₈₃ N ₇ O ₁₄ : 1053.60. Found (m / z): 1054.9 (M + H) ⁺ .

Example 63

Synthesis of [Val] ³ -aplidine (SVPL1)

Follow the procedure developed for the synthesis of SAPL1 based on SVPL2 (10 mg, 11 μmol) and F1 (10.5 mg, 57 μmol). The title compound (8 mg, 69%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, isotrate ratio ACN / H ₂ O 85:15 (flow: 7 ml / min, 250 × 21 mm, 270 nm, t _R = 10.9 and 12.3 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.02 (m, 24H), 1.13-1.38 (m, 9H), 1.55 (m, 2H), 1.67 -1.81 (m, 4H), 1.95-2.02 (m, 3H), 2.10-2.17 (m, 2H), 2.26-2.39 (m, 2H), 2 56 (s, 3H), 2.57 (s, 3H), 2.58 (s, 3H), 2.74-2.92 (m, 1H), 3.10 (s, 3H), 3, 15 (s, 3H), 3.20 (m, 1H), 3.36 (dd, J ₁ = 4.4, J ₂ = 14.2, 1H), 3.49-3.72 (m, 5H ), 3.79 (s, 3H), 3.97-4.13 (m, 2H), 4.38 (dd, J ₁ = 4.6, J ₂ = 14.2, 1H), 4.49 (m, 1H), 4.60 (m, 1H), 4.68-4.81 (m, 2H), 5.11 (m, 1H), 5.26-5.30 (m, 1H), 5.33-5.40 (m, 1H), 6.84 (d, J = 7.8, 2H), 7.08 (d, J = 8.3, 2H), 7.36-7.52 (m, 2H), 7.48 (d, J = 9.6, 1H), 7.61 (d, J = 6.8, 1H).

ESI-MC Calculate. for C ₅₄ H ₈₄ N ₈ O ₁₃ 1052.62. Found (m / z): 1053.6 (M + H) ⁺ .

Example 64

Synthesis of [Hiv] ³ - [Isobutyryl] ^8- didemnin A (8ISHPL1)

To a solution of SHPL2 (10 mg, 11.2 μmol) in DCM (200 μl) at 0 ° C in an Ar atmosphere, DIPEA (3 μl, 16.8 μmol) and isobutyryl chloride (1.4 μl, 13.4 μmol) were added. After 3 hours at 22 ° C, DCM (3 ml) was added and the mixture was washed successively with aq. HCl (2 ml, 0.1 N) aq. NaHCO ₃ (2 ml, sat.) And brine (2 ml), dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. Purification of the residue by HPLC (HyperPrep PEP 100 C18, isotrate ratio ACN / H ₂ O 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 19 min) gives the title compound (10 mg, 94%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (24H, m), 1.13-1.65 (18H, m), 1.72-2.58 (6H, m) 2.56 (s, 3H), 2.89 (s, 3H), 2.92 (m, 2H), 3.13 (m, 2H), 3.36 (dd, J ₁ = 4.6, J ₂ = 15.6, 1H), 3.54-3.73 (m, 3H), 3.78 (s, 3H), 3.91 (m, 2H), 4.40 (m, 1H), 4.60 (m, 1H), 4.89 (m, 1H), 4.99 (d, J = 5.3, 1H), 5.03 (m, 1H), 5.20 (m, 1H) 6.73 (d, J = 9.3, 1H); 6.84 (d, J = 9.6; 2H); 7.08 (d, J = 9.6; 2H); 7.55 ( d, J = 8.6, 1H); 7.82 (d, J = 11, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.78, 14.63, 15.69, 17.92, 19.03, 19.31, 19.66, 21.03, 22.29, 23, 19, 23.85, 24.71, 25.14, 27.27, 28.22, 30.39, 30.51, 31.15, 33.80, 34.25, 35.61, 38.85, 39.63, 46.98, 48.51, 53.29, 53.65, 55.49, 56.01, 56.35, 57.26, 66.07, 69.07, 70.94, 79, 33, 114.31, 130.08, 130.60, 158.84, 168.79, 169.75, 170.34, 170.68, 171.36, 171.73, 174.02, 179.84.

ESI-MC Calculate. for C ₅₀ H ₈₀ N ₆ O ₁₂ : 956.58. Found (m / z): 957.5 (M + H) ⁺ .

Example 65

Synthesis of [Val] ³ - [Isobutyryl] ^8- didemnin A (8ISVPL1)

Follow the procedure developed for the synthesis of 8ISHPL1 based on SVPL2 (20 mg, 22.6 μmol). The title compound (19 mg, 88%) was obtained after purification by HPLC (HyperPrep PEP 100 C18, AFM / H ₂ O ratio 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 19 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.81-1.02 (m, 24H), 1.14-1-38 (m, 5H), 1.15 (d, J = 6.6, 3H), 1.19 (d, J = 6.6, 3H), 1.38-1.80 (m, 7H), 1.80-2.40 (m, 6H), 2.57 (s, 3H), 2.58-2.64 (m, 1H), 2.85-2.92 (m, 1H), 2.93 (s, 3H), 3.16 (dd, J ₁ = 10.5 , J ₂ = 14.4, 1H), 3.36 (dd, J ₁ = 4.5, J ₂ = 14.4, 1H), 3.39 (br s, 1H), 3.56 (dd , 1H, J = 4.5, 10.8), 3.59-3.72 (m, 3H), 3.78 (s, 3H), 4.01 (td, J ₁ = 3.3, J ₂ = 10.2, 1H), 4.39-4.47 (m, 2H), 4.58 (dd, J ₁ = 5.7, J ₂ = 7.5, 1H), 4.79 (t , J = 9.9, 1H), 5.03-5.14 (m, 2H), 6.84 (d, J = 8.4, 2H), 7.04 (d, J = 7.8, 1H), 7.08 (d, J = 8.4, 1H), 7.36 (d, J = 9.0, 1H), 7.45 (d, J = 10.2, 1H), 7, 51 (d, J = 10.2, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 180.24, 175.10, 173.18, 171.14, 170.72, 170.42, 169.38, 168.73, 158.87, 130, 60, 130.02, 114.33, 71.31, 70.50, 66.11, 59.47, 57.42, 56.56, 55.51, 54.93, 53.84, 48.83, 47.13, 41.94, 38.94, 35.67, 34.17, 33.63, 31.32, 31.10, 29.96, 28.32, 27.25, 25.30, 25, 05, 24.82, 24.01, 23.22, 21.42, 19.88, 19.72, 11.25, 18.51, 15.60, 14.32, 11.96.

ESI-MC Calculate. for C ₅₀ H ₈₁ N ₇ O ₁₁ : 955.60. Found 956.8 (M + H) ⁺ .

Example 66

Synthesis of [Hiv] ³ - [butyryl] ^8- didemnin A (8BSHPL1)

Follow the procedure developed for the synthesis of 8ISHPL1 based on SHPL2 (10 mg, 11.2 μmol). The title compound (9 mg, 84%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, isotrate ratio ACN / H ₂ O 85:15 (flow rate: 7 ml / min, 250 × 21 mm, 270 nm, t _R = 18.6 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 24H), 1.11-1.72 (m, 18H), 1.75-2.51 (m, 6H) 2.56 (s, 3H), 2.84 (s, 3H), 2.92 (m, 2H), 3.15 (m, 2H), 3.35 [dd, J ₁ = 5.0, J ₂ = 15.6, 1H), 3.54-3.77 (m, 3H), 3.79 (s, 3H), 3.91 (m, 2H), 4.40 (m, 1H), 4.60 (m, 1H), 4.89 (m, 1H), 4.98 (d, J = 6.0, 1H), 5.04 (m, 1H), 5.20 (m, 1H) 6.78 (d, J ₁ = 9.6, 1H), 6.84 (d, J ₁ = 9.6, 2H), 7.08 (d, J ₁ = 9.6, 2H), 7 54 (d, J ₁ = 9.6, 1H); 7.82 (d, J ₁ = 10.6, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.82, 14.15, 14.55, 15.72, 17.96, 18.67, 19.01, 21.05, 22.37, 23, 13, 23.84, 24.71, 25.14, 27.30, 28.21, 29.92, 30.38, 30.63, 33.78, 34.25, 35.69, 35.97, 38.85, 39.61, 46.98, 48.52, 53.27, 55.49, 55.99, 56.25, 57.27, 66.07, 69.07, 70.96, 79, 40, 114.31, 130.08, 130.59, 158.85, 168.78, 169.73, 170.42, 170.64, 171.36, 171.65, 174.07, 175.79.

ESI-MC Calculate. for C ₅₀ H ₈₀ N ₅ O ₁₂ : 956.60. Found (m / z): 957.8 (M + H) ⁺ .

Example 67

Synthesis of [Hiv] ³ - [Hexanoyl] ^8- didemnin A (8BSHPL1)

Following the procedure developed for the synthesis of 8ISHPL1, starting from SHPL2 (10 mg, 11.2 μmol), the title compound (9 mg, 82%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, iso ACN ratio / H ₂ O 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 27.8 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 24H), 1.11-1.72 (m, 22H), 1.80-2.51 (m, 6H) 2.56 (s, 3H), 2.84 (s, 3H), 2.93 (m, 2H), 3.14 (m, 2H), 3.35 (dd, J ₁ = 4.4, J ₂ = 14.1, 1H), 3.54-3.76 (m, 3H), 3.79 (s, 3H), 3.91 (m, 2H), 4.41 (m, 1H), 4.60 (m, 1H), 4.88 (m, 1H), 4.98 (d, J ₁ = 5.3, 1H), 5.03 (m, 1H), 5.19 {m, 1H ), 6.76 (d, J ₁ = 8.7, 1H), 6.84 (d, J ₁ = 8.7, 2H), 7.08 (d, J ₁ = 8.7, 2H), 7.51 (d, J ₁ = 8.8, 1H); 7.81 (d, J ₁ = 9.7, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.59, 13.94, 14.34, 15.49, 17.73, 18.77, 20.81, 22.14, 22.46, 22, 90, 23.60, 24.47, 24.69, 24.90, 27.08, 27.97, 30.14, 30.40, 31.54, 33.53, 33.76, 34.02, 35.47, 38.62, 39.40, 46.76, 48.28, 53.04, 55.26, 55.78, 55.90, 57.04, 65.84, 68.82, 70, 72, 79.17, 114.07, 129.84, 130.36, 158.61, 168.54, 169.51, 170.15, 170.41, 171.12, 171.42, 173.86, 175.75.

ESI-MC Calculate. for C ₅₂ H ₈₄ N ₆ O ₁₂ : 984.61. Found (m / z): 985.8 (M + H) ⁺ .

Example 68

Synthesis of Isobutyryl-Pro-OBn

To a solution of H-Pro-OBn · HCl (500 mg, 2.07 mmol) in DCM (10 ml) at 0 ° C was added NMR (680 μl, 6.21 mmol) in argon atmosphere. After 10 minutes, isobutyryl chloride (240 μl, 2.27 mmol) was added, the reaction mixture was allowed to warm to 20 ° C and stirred for 5 hours. The mixture was filtered and the filtrate was washed successively with water. HCl (15 ml, 1 N), water. NaHCO ₃ (10 ml, sat.) And brine (10 ml), then dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to obtain 560 mg (98%) of the title compound. Rf = 0.42 (hex.: EtOAc 1: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.20-1.40 (2d, 6H), 1.90-2.35 (m, 4H), 2.35 (q, 1H), 3.40 -3.80 (m, 2H), 4.30 (m, 1H), 5.20 (m, 2H), 7.40 (m, 5H).

Example 69

Synthesis of Isobutyryl-Pro-OH

To a solution of isobutyryl-Pro-OBn (430 mg, 1.56 mmol) in degassed MeOH was added Pd / C (10%) (43 mg, 10% m / m) and then sequentially purged with a strong stream of Ar and bubbled hydrogen. The mixture was stirred under H _{2 for} 14 hours and then degassed and filtered. The solution was concentrated and the residue was crystallized with MTBE / hex. To give 140 mg (48%) of the title compound as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.20 (m, 6H), 1.90-2.10 (m, 3H), 2.50 (m, 1H), 2.70 (q, 1H ), 3.40-3.70 (m, 2H), 4.60 (dd, 1H).

ESI-MC Calculate. for C ₉ H ₁₅ NO ₃ : 185.11. Found (m / z): 186.1 (M + H) ⁺ .

Example 70

Synthesis of [Hiv] ³ - [Isobutyryl] ⁹ -aplidine (9ISHPL1)

To a solution of isobutyryl-Pro-OH (10 mg, 54 μmol) in DCM (150 μl) at 0 ° C was added DIPCDI (5 μl, 32 μmol). Stirring was continued for 60 minutes and then the mixture was transferred to a flask containing SHPL2 (10 mg, 11.2 mol) in DCM (150 L). After 4 hours at 2-4 ° C., the mixture was diluted with DCM (2 ml) and washed successively with aq. HCl (1 ml, 0.1 N) aq. NaHCO ₃ (1 ml, sat.) And brine (1 ml), the organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. The residue was purified by HPLC (HyperPrep PEP 100 C18, isotrate ratio ACN / H ₂ O 85:15 (flow: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 13 and 14 min), which gives 9ISHPL1 (9 mg, 72%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 24H), 1.12-1.50 (m, 18H), 1.52-2.70 (m, 10H) 2.56 (s, 3H), 3.00-3.44 (m, 4H), 3.08 (s, 3H), 3.55-3.72 (m, 5H) 3.78 (s, 3H), 4.00 (m, 3H), 4.26 (m, 1H), 4.62 (m, 2H), 4.86 (m, 1H), 5.02 (d, J = 5.3 , 1H), 5.30 (m, 2H), 6.84 (d, J = 9.0, 2H), 7.07 (d, J = 9.0, 2H), 7.29 (d, J = 11.0, 1H); 7.80 (d, J = 9.0, 1H); 7.88 (d, J = 11, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 12.04, 14.44, 16.91, 17.94, 18.96, 19.03, 19.15, 21.07, 21.59, 23, 87, 24.10, 24.89, 25.08, 26.04, 27.54, 28.21, 28.87, 30.33, 31.61, 32.64, 33.77, 34.24, 36.19, 39.07, 39.33, 39.81, 46.87, 47.52, 48.45, 54.74, 55.49, 56.20, 57.08, 58.50, 66, 41, 69.13, 71.48, 79.16, 114.27, 130.30, 130.59, 158.80, 168.55, 169.88, 170.86, 171.06, 171.22, 173.63, 174.92, 176.19.

ESI-MC Calculate. for C ₅₅ H ₈₇ N ₇ O ₁₃ : 1053.64. Found (m / z): 1054.6 (M + H) ⁺ .

Example 71

Synthesis of [Val] ³ - [Isobutyryl] ⁹ -aplidine (9ISVPL1)

Following the procedure developed for the synthesis of 9ISHPL1, starting from SVPL2 (10 mg, 11.2 μmol), the title compound (9 mg, 77%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, iso ACN ratio / H ₂ O 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 15.3 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.01 (m, 24H), 1.14-1.37 (m, 12H), 1.148-2.35 (m, 8H), 2 55 (s, 3H), 2.57-2.68 (m, 1H), 2.77-2.83 (m, 1H), 3.12 (s, 3H), 3.19-3.23 (m, 1H), 3.35-3.41 (m, 2H), 3.52-3.56 (m, 1H), 3.58-3.70 (m, 3H), 3.78 (s , 3H), 4.03-4.13 (m, 1H), 4.33-4.35 (m, 1H), 4.44-4.48 (m, 1H), 4.55-4.66 (m, 2H), 4.70-4.83 (m, 1H), 5.35-5.39 (m, 2H), 6.82 (d, J = 8.4, 2H), 7.06 (d, J = 8.4, 2H), 7.25 (br s, 2H), 7.37 (d, 10.5, 1H), 7.46 (d, J = 8.7, 1H) 7.60 (d, J = 9.3, 1H); 8.07 (br s, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 12.10, 14.44, 16.84, 18.63, 18.95, 19.92, 21.31, 21.61, 23.91, 24, 05, 24.86, 25.00, 25.30, 26.06, 27.41, 28.33, 28.93, 30.00, 31.76, 32.65, 33.59, 31.16, 36.21, 39.17, 41.71, 42.19, 47.08, 47.57, 48.88, 54.37, 54.65, 55.50, 56.15, 57.33, 58, 70, 59.33, 66.45, 70.92, 71.54, 114.29, 130.28, 130.59, 158.82, 168.41, 170.08, 170.52, 170.69, 171.03, 172.65, 173.80, 175.68, 176.42.

ESI-MC Calculate. for C ₅₅ H ₈₈ N ₈ O ₁₃ : 1052.65. Found (m / z): 1053.8 (M + H) ⁺ .

Example 72

Synthesis of Z-NVa-Pro-OMe

Following the procedure developed for the synthesis of F2, from Z-NVa-OH (261 mg, 1.04 mmol), H-Pro-OMe-HCl (156.6 mg, 0.94 mmol), the title compound (315 mg , 87%) are obtained as a colorless oil after purification by LC-silica (hex. - EtOAc, gradient 3: 1 - 1: 1). R _t = 0.42 (hex. - EtOAc 1: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.97 (t, J = 6.9, 2H), 1.44 (sext., J = 7.5, 2H), 1.53-1.65 (m, 1H), 1.68-1.77 (m, 1H), 1.82-2.09 (m, 3H), 2.17-2.24 (m, 1H), 3.43-3 80 (m, 2H), 3.71 (s, 3H), 4.45-4.55 (m, 2H), 5.07 (s, 2H), 5.51 (d, J = 8.4 , 1H); 7.32-7.35 (m, 5H).

Example 73

Synthesis of NVa-Pro-OH

To a solution of Z-NVa-Pro-OMe (36 mg, 99 μmol) in a mixture of THF and MeOH (130 μl: 130 μl) at 0 ° C was added aq. LiOH (130 μl, 15% m / m). After 6 hours of stirring, the reaction mixture was partitioned between H ₂ O (3 ml) and diethyl ether (3 × 2 ml). The organic phase is then extracted with NaHCO ₃ (3 × 2 ml). The combined aqueous phases neutralize (pH 5) aq. HCl (0.1 N) and extracted with Et ₂ O (3 × 3 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered and concentrated to give the title compound (36 mg, quant.) As a colorless oil.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.96 (t, J = 6.9, 2H), 1.41 (sext., J = 7.4, 2H), 1.53-1.65 -1.77 (m, 2H), 1.82-2.10 (m, 3H), 2.17-2.24 (m, 1H), 3.52-3.81 (m, 2H), 4 , 45-4.58 (m, 2H), 5.07 (br s, 2H), 5.81 (d, J = 8.4, 1H), 7.30-7.35 (m, 5H) 7.41 (br s, 1H).

ESI-MC Calculate. for C ₁₈ H ₂₄ N ₂ O ₅ : 348.17. Found (m / z): 349.2 (M + H) ⁺ .

Example 74

Synthesis of [ZNVa-Pro] 9-aplidine (9NVSAPL2)

Following the procedure developed for the synthesis of SAPL1, starting from SAPL2 (18 mg, 19 μmol) and Z-NVa-Pro-OH (34 mg, 97 μmol), the title compound (16 mg, 66%) was obtained as a white solid substances after purification by HPLC (HyperPrep PEP 100 C18, isotrate ratio ACN / H ₂ O 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 29 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.84-0.96 (m, 27H), 1.12-1.85 (m, 19H), 2.00-2.25 (m, 7H) 2.30-2.40 (m, 1H), 2.54 (s, 3H), 2.62 (dd, J ₁ = 10.5, J ₂ = 17.7, 1H), 2.93 ( d, 4.2, 1H), 3.14 (s, 3H), 3.14-3.20 (m, 1H), 3.28-3.34 (m, 2H), 3.50-3, 67 (m, 4H), 3.77-3.80 (m, 1H), 3.79 (s, 3H), 3.82-3.91 (m, 1H), 4.00-4.17 ( m, 2H), 4.27 (dd, J ₁ = 6.3, J ₂ = 13.2, 1H), 4.43-4.51 (m, 2H), 4-58-4.63 (m , 1H), 4.69-4.75 (m, 1H), 4.77-4.82 (m, 1H), 5.07 (d, 1H, J = 12.9, 2H), 5.13 (d, J = 12.9, 1H), 5.32-5.41 (m, 2H), 6.07 (d, J = 8.7, 1H), 6.83 (d, J = 8, 4, 2H), 7.06 (d, J = 8.4, 2H), 7.17 (d, J = 9, 9, 1H), 7.32 (m, 5H), 7.83 (d, J = 9.0, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.68, 13.74, 14.56, 15.24, 16.39, 16.85, 18.61, 20.94, 21.26, 23, 34, 23.70, 24.76, 24.90, 25.02, 26.00, 27.17, 27.81, 28.54, 31.29, 31.40, 33.30, 33.85, 33.86, 36.19, 38.62, 38.84, 41.31, 46.91, 47.21, 49.38, 49.49, 52.50, 54.96, 55.24, 55, 25, 56.50, 57.17, 57.96, 62.53, 66.40, 67.93, 70.64, 81.37, 114.04, 127.89, 127.77, 128.32, 129.97, 130.29, 136.84, 156.72, 158.55, 168.48, 169.36, 169.58, 170.52, 171.27, 171.71, 172.54, 173, 22, 205.08.

ESI-MC Calculate. for C ₆₇ H ₁₀₀ N ₈ O ₁₆ : 1277.3, Found (m / z): 1273.7 (M + H) ⁺ .

Example 75

Synthesis of [Hiv] ³ - [Z-NVa-Pro] ⁹ -aplidine (9NVSHPL2)

Following the procedure developed for the synthesis of SAPL1, starting from SHPL2 (10 mg, 11.2 mmol) and Z-NVa-Pro-OH (20 mg, 56 μmol), the title compound (8 mg, 60%) was obtained as white solid after purification by HPLC (HyperPrep PEP 100 C18, isotrate ratio ACN / H ₂ O 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 26.7 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 24H), 1.12-1.80 (m, 22H), 1.82-2.35 (m, 6H) 2.42 (m, 1H), 2.56 (s, 3H), 2.96-3.38 (m, 4H), 3.10 (s, 3H), 3.48-3.72 (m 5H), 3.78 (s, 3H), 3.88 (m, 1H), 4.01 (m, 1H), 4.18 (m, 1H), 4.47 (m, 1H), 4 68 (m, 2H), 4.87 (m, 1H), 5.02 (d, J ₁ = 5.3, 1H), 5.08 (m, 2H), 5.28 (m, 1H) 5.42 (m, 1H), 6.10 (d, J = 8.3, 1H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8, 3, 2H), 7.31 (m, 6H), 7.72 (d, J = 4.3, 1H); 7.78 (d, J = 8.7, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 12.06, 13.88, 14.22, 16.85, 17.76, 18.89, 19.18, 21.12, 21.47, 23, 72, 23.98, 25.08, 26.25, 27.57, 28.15, 28.79, 30.32, 31.61, 33.45, 33.72, 34.07, 35.95, 38.92, 39.59, 39.90, 46.87, 47.44, 48.46, 52.76, 55.21, 55.49, 56.77, 57.17, 58.37, 66, 41, 66.65, 69.22, 71.14, 79.07, 114.24, 127.87, 128.00, 128.56, 130.25, 130.55, 157.00, 158.79, 168.82, 169.86, 170.36, 170.58, 170.77, 171.30, 171.86, 173.28, 174.94.

ESI-MC Calculate. for C ₆₄ H ₉₆ N ₈ O ₁₅ : 1216.7. Found m / z: 1217.5 (M + H) ⁺ .

Example 76

Synthesis of [NVa-Pro] ⁹ -aplidine (9NVSAPL1)

A degassed mixture of 9NVSAPL2 (10 mg, 7.8 μmol) and Pd / C (10%, 5 mg) in IPA: H ₂ O (0.2 ml: 0.1 ml) is saturated (and held at 1 atm) with hydrogen gas with stirring for 14 hours. Then the mixture was filtered (Teflon, 0.45 μm) and concentrated in vacuo to give the title compound (8.8 mg, quant.) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.85-0.95 (m, 27H), 1.18-1.51 (m, 18H), 1.50-2.45 (m, 7H) 2.59-2.83 (m, 1H), 2.57 (s, 3H), 2.57-2.80 (m, 3H), 2.81-2.95 (m, 1H), 3 14 (s, 3H), 3.15-3.40 (m, 3H), 3.52-3.79 (m, 4H), 3.79 (s, 3H), 4.45-4.52 (m, 1H), 4.61-4.65 (m, 1H), 4.70-4.85 (m, 2H), 5.17 (d, J = 3.3, 1H), 5.36 -5.39 (m, 2H), 6.84 (d, J = 8.1, 2H), 7.86 (d, J = 8.7, 2H), 7.19 (d, J = 10, 2, 1H), 7.82 (d, J = 9.0, 1H), 7.80-7.85 (m, 1H).

ESI-MC Calculate. for C ₅₉ H ₄₀ N ₈ O ₁₄ : 1138.69. Found (m / z): 1193.7 (M + H) ⁺ .

Example 77

Synthesis of [Hiv] ³ - [NVa-Pro] ⁹ -aplidine (9NVSHPL1)

Following the procedure developed for the synthesis of 9NVSAPL1, starting from 9NVSHPL2 (10 mg, 8.2 μmol), the title compound (8 mg, quant.) Was obtained as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 30H), 1.12-1.85 (m, 22H), 1.92-2.35 (m, 6H) 2.42 (m, 1H), 2.56 (s, 3H), 3.10-3.45 (m, 4H), 3.10 (s, 3H), 3.50-3.70 (m 5H), 3.78 (s, 3H), 3.82 (m, 1H), 4.01 (m, 1H), 4.26 (m, 1H), 4.65 (m, 1H), 4 64 (m, 1H), 4.88 (m, 1H), 5.02 (d, J = 5.3, 1H), 5.32 (m, 2H), 6.84 (d, J = 8 , 3, 2H), 7.07 (d, J = 8.3, 2H), 7.38 (d, J = 8.7, 1H), 7.60 (d, J = 4.3, 1H) 7.80 (d, J = 9.3, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 12.03, 14.13, 14.30, 16.80, 17.81, 18.72, 19.15, 21.09, 21.55, 23, 73, 23.96, 25.07, 26.16, 27.55, 28.17, 28.72, 29.91, 30.33, 31.52, 33.80, 34.17, 36.00, 38.94, 39.51, 39.84, 46.88, 47.51, 48.45, 55.01, 55.49, 56.91, 57.15, 58.10, 66.37, 69, 17, 71.10, 79.12, 114.29, 130.21, 130.57, 158.82, 163.66, 168.91, 169.86, 170.38, 170.75, 170.82, 171.30, 173.22, 174.79.

ESI-MC Calculate. for C ₅₆ H ₉₀ N ₈ O ₁₃ : 1082.66. Found m / z: 1083.7 (M + H) ⁺ .

Example 78

Synthesis of [Hiv] ³ - [L-Lac (OTBDMS)] ⁹ -aplidine [9LSHPL2 (L)]

Following the procedure developed for the synthesis of SAPL1, starting from SHPL2 (10 mg, 11.2 μmol) and (L) -Lac (OTBDMS) -Pro-OH (17 mg, 56 μmol), the title compound (9 mg, 68 %) is obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, gradient ACN / H ₂ O 85:15 - 100: 0 in 10 min (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 30.1 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.08 (s, 3H), 0.10 (s, 3H), 0.82-1.10 (m, 24H), 1.11-1.72 (m, 18H), 1.75-2.51 (m, 6H), 2.41 (m, 1H), 2.56 (s, 3H), 3.00-3.40 (m, 5H), 3.11 (s, 3H), 3.53-3.82 (m, 3H), 3.79 (s, 3H), 3.91 (m, 2H), 4.02 (m, 1H), 4 27 (m, 1H), 4.50 (m, 1H), 4.63 (m, 2H), 4.87 (m, 1H), 5.01 (d, J = 4.8, 1H), 5.27 (m, 2H), 6.84 (d, J = 8.7, 2H), 7.07 (d, J 8.7, 2H), 7.29 (d, J = 9.7, 1H), 7.63 (d, J = 5.8, 1H), 7.88 (d, J = 9.7, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ -4.26, -4.12, 12.04, 14.40, 16.91, 17.97, 19.14, 20.60, 21.11, 21.66, 23.82, 24.11, 24.96, 25.08, 26.11, 26.37, 27.53, 28.18, 28.37, 30.33, 31.69, 33, 75, 34.19, 36.23, 39.00, 39.36, 39.81, 46.87, 47.64, 48.45, 54.91, 55.37, 55.48, 56.81, 57.08, 58.37, 66.38, 69.14, 69.89, 71.42, 79.19, 82.66, 114.23, 130.36, 130.59, 158.77, 168, 43, 469.86, 170.72, 171.01, 171.21, 172.03, 173.62, 174.91.

ESI-MC Calculate. for C ₆₀ H ₉₉ N ₇ O ₁₄ Si: 1169.7. Found m / z: 1170.9 (M + H) ⁺ .

Example 79

Synthesis of [Hiv] ³ - [D-Lac (OTBDMS)] ^9- aplidine [9LSHPL2 (D)]

Following the procedure developed for the synthesis of SAPL1, starting from SHPL2 (10 mg, 11.2 μmol) and (D) -Lac (OTBDMS) -Pro-OH (17 mg, 56 μmol), the title compound (9 mg, 68 %) is obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, gradient ACN / H ₂ O 85:15 - 100: 0 in 10 min (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 30.4 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.03 (m, 3H), 0.06 (m, 3H), 0.87 (s, 9H), 0.82-1.10 (m, 24H ), 1.11-1.72 (m, 18H), 1.75-2.30 (m, 6H), 2.41 (m, 1H), 2.56 (s, 3H), 3.00- 3.40 (m, 5H), 3.06 (s, 3H), 3.56 (m, 1H), 3.65 (m, 2H), 3.78 (s, 3H), 3.90 (m , 1H), 4.01 (m, 1H), 4.17 (m, 1H), 4.25 (m, 1H), 4.39 (m, 1H), 4.61 (m, 2H), 4 86 (m, 1H), 5.00 (d, J = 4.8, 1H), 5.25 (m, 2H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8.3, 2H), 7.29 (d, J = 9.7, 1H), 7.74 (d, J = 5.3, 1H), 7.87 (d, J = 9.7, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ -4.87, -4.84, 12.04, 14.38, 16.89, 17.94, 19.14, 20.24, 21.08, 21.60, 23.85, 24.12, 24.90, 25.06, 25.13, 26.01, 26.53, 27.53, 27.83, 28.20, 30.33, 31, 51, 33.76, 34.20, 36.16, 38.99, 39.35, 39.82, 46.87, 47.02, 48.46, 54.79, 55.42, 55.48, 57.08, 57.18, 58.28, 66.39, 69.11, 71.43, 72.61, 79.18, 114.25, 130.29, 130.59, 158.79, 168, 45, 169.86, 170.77, 170.81, 170.97, 171.20, 172.53, 173.67, 174.84.

ESI-MC Calculate. for C ₆₀ H ₉₉ N ₇ O ₁₄ Si: 1169.7. Found m / z: 1170.8 (M + H) ⁺ .

Example 80

Synthesis of [Val] ³ - [L-Lac (OTBDMS)] ^9- aplidine [9LSVPL2 (L)]

Following the procedure developed for the synthesis of SAPL1, starting from SVPL2 (10 mg, 11.2 μmol) and (L) -Lac (OTBDMS) -Pro-OH (17 mg, 56 μmol), the title compound (9 mg, 68 %) is obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, iso-ratio of ACN / H ₂ O 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 17.8 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.14 (s, 6H), 0.71-1.06 (m, 27H), 1.10-1.42 (m, UN), 1.43 -1.84 (m, 8H), 1.85-2.40 (m, 11H), 2.57 (s, 3H), 2.80 (d, J = 14.7, 1H), 3.15 (s, 3H), 3.15-3.23 (m, 1H), 3.33-3.42 (m, 2H), 3.54 (dd, J ₁ = 4.2, J ₂ = 10, 8, 1H), 3.58-3.69 (m, 3H), 3.79 (s, 3H), 3.88-3.90 (m, 1H), 4.05-4.12 (br. t, 1H), 4.32 (br s, 1H), 4.43-4.68 (m, 4H), 4.77 (t, J = 10.5, 1H), 5.31-5, 35 (m, 2H), 6.83 (d, J = 8.4, 2H), 7.08 (d, J = 8.7, 2H), 7.39 (d, J = 9.9, 1H ), 7.45 (d, J = 9.0, 1H), 7.60 (d, J = 10.5, 1H), 7.83 (d, J = 4.5, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ -4.67, -4.49, 11.84, 14.13, 16.67, 18.19, 18.41, 19.68, 20.48, 21.15, 21.40, 23.61, 23.92, 24.72, 24.80, 25.06, 25.88, 26.24, 27.17, 28.04, 28.13, 29, 78, 31.57, 33.33, 33.81, 35.99, 38.84, 41.56, 41.95, 46.83, 47.52, 48.68, 54.09, 54.64, 55.26, 56.63, 57.13, 58.36, 59.12, 66.20, 70.10, 70.57, 71.27, 77.20, 114.01, 130.13, 130, 36, 158.55, 168.09, 169.76, 170.07, 170.48, 170.77, 172.21, 172.33, 173.58, 175.45.

ESI-MC Calculate. for C ₅₉ H ₄₀ N ₈ O ₁₄ : 1168.72. Found (m / z): 1169.8 (M + H) ⁺ .

Example 81

Synthesis of [Hiv] ³ - [L-Lac] ⁹ -aplidine [9LSHPL1 (L)]: tamandarin A

To a solution of 9LSHPL2 (L) (16 mg, 14 μmol) in THF (500 μl, anhydrous) at 0 ° C in an Ar atmosphere was added TBAF (50 μl, 1M in THF). After 1 h at 22 ° C, the mixture was concentrated in vacuo and the crude product was purified by flash LC (silica gel, deg. DCM: MeOH 1% -5%) to obtain the title compound (813 mg, 88%) as a white solid.

Experimental data published: Fenical, W. et al., J. Org. Chem. 2000, 65, 782-792.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-0.96 (m, 18H), 1.02 (d, J = 3.4, 3H), 1.04 (d, J = 3, 4, 3H), 1.14-2.28 (m, 14H), 1.24 (s, 3H), 1.34 (d, J = 6.8, 3H), 1.43 (d, J = 6.8, 3H), 2.44 (dd, J ₁ = 7.8, J ₂ = 17.1, 1H), 2.58 (s, 3H), 3.00 (br s, 1H), 3.10 (s, 3H), 3.14-3.31 (m, 2H), 3.37-3.43 (m, 2H), 3.56-3.72 (m, 5H), 3, 79 (s, 3H), 3.90 (t, J = 7.8, 1H), 4.02 (dt, J ₁ = 3.4, J ₂ = 9.8, 1H), 4.25 (d , J = 3.9, 1H), 4.30 (t, J = 6.8, 1H), 4.37 (dd, J ₁ = 7.3, J2 = 8.3, 1H), 4.65 (m, 1H), 4.71 (t, J = 7.4, 1H), 4.87 (t, J = 11.2, 1H), 5.03 (d, J = 4.9, 1H) 5.29 (dd, J ₁ = 3.4, J ₂ = 11.7, 1H), 5.42 (m, 1H), 6.83 (d, J = 8.3, 2H), 7, 07 (d, J = 8.3, 2H], 7.34 (d, J = 9.8, 1H), 7.48 (d, J = 5.4, 1H), 7.76 (d, J = 9.8, 1H).

ESI-MC Calculate. for C ₅₄ H ₈₅ N ₇ O ₁₄ : 1055.6. Found: 1056.7 (M + H) ⁺ .

Example 82

Synthesis of [Hiv] ³ - [D-Lac] ⁹ -aplidine [9LSHPL1 (D)]

Following the procedure developed for the synthesis of 9LSHPL1 (L), starting from 9LSHPL2 (D) (20 mg, 17 μmol) and TBAF (50 μl, 1M in THF), the title compound (14 mg, 78%) is obtained as white solid, after purification by flash LC (silica gel, deg. DCM: MeOH 1% -5%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.78-1.08 (m, 18H), 1.02 (d, J = 3.9, 3H), 1.04 (d, J = 3, 4, 3H), 1.10-2.36 (m, 14H), 1.20 (d, J = 6.3, 3H), 1.34 (d, J = 6.3, 3H), 1, 37 (d, J = 6.3, 3H), 2.38-2.50 (dd, J ₁ = 7.8, J ₂ = 17.5, 1H), 2.56 (s, 3H), 3 10 (s, 3H), 3.13-3.20 (m, 1H), 3.22-3.28 (m, 1H), 3.37-3.42 (dd, J ₁ = 3.9 , J ₂ = 4.3, 1H), 4.61-4.68 (m, 3H), 3.69-3.76 (m, 3H), 3.77 (m, 1H), 3.78 ( s, 3H), 3.90 (t, J = 7.8, 1H), 3.97-4.07 (m, 1H), 4.26 (m, 1H), 4.41 (q, J = 6.3, 1H), 4.63 (m, 1H), 4.71 (m, 1H), 4.86 (t, J = 10.7, 1H), 5.01 (d, J = 4, 8, 1H), 5.21-5.37 (m, 2H), 6.83 (d, J = 8.3, 2H), 7.06 (d, J = 8.3, 2H), 7, 41 (m, 2H), 7.77 (d, J = 9.2, 1H).

ESI-MC Calculate. for C ₅₄ H ₈₅ N ₇ O ₁₄ : 1055.62. Found: 1056.6 (M + H) ⁺ .

Example 83

Synthesis of [Val] ³ - [L-Lac] ⁹ -aplidine [9LSVPL1 (L)]

Following the procedure developed for the synthesis of 9LSHPL1, starting from 9LSHPL2 (D) (5 mg, 4.3 μmol), the title compound (4 mg, 88%) is obtained as a white solid, after purification by flash LC (silica gel, grad DCM: MeOH 1% -5%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.02 (m, 18H), 1.16-1.42 [i, 4H), 1.32 (d, J = 3.0, 3H), 1.40 (d, J = 6.6, 3H), 1.56-1.83 (m, 8H), 1.95-2.34 (m, 11H), 2.58 (s, 3H), 2.84 (d, J = 14.7, 1H), 3.15 (s, 3H), 3.15-3.23 (m, 1H), 3.36-3.42 (m, 1H), 3.55 (dd, J ₁ = 9.0, J ₂ = 10.5, 2H), 3.64-3.66 (m, 3H), 3.95 (dd, J ₁ = 3, 3, J ₂ = 9.9, 1H), 4.08 (td, J ₁ = 7.5, J ₂ = 17.1, 1H), 4.32 (br s, 1H), 4.41 ( dd, J ₁ = 6.6, J ₂ = 9.9, 1H), 4.48 (dd, J ₁ = 5.1, J ₂ = 10.5, 1H), 4.61 (dd, J ₁ = 6.0, J ₂ = 6.6, 1H), 4.69-4.80 (m, 2H), 5.29-5.35 (m, 1H), 5.57 (m, 1H), 6.84 (d, J = 8.1, 2H), 7.08 (d, J = 8.7, 2H), 7.37 (d, J = 3.9, 1H), 7.40 (d , J = 5.4, 1H), 7.60 (d, J = 10.8, 1H), 7.72 (d, J = 3.9, 1H).

ESI-MC Calculate. for C ₅₄ H ₈₆ N ₈ O ₁₃ : 1054.63. Found: 1055.8 (M + H) ⁺ .

Synthesis of spiro [4,4] nonane fragment:

Example 84

Synthesis of N - [(2R) -2-allyl-N- (tert-butoxycarbonyl) prolyl] D-leucine (9)

To a cooled (0 ° C) solution of 8 (1.53 g, 6 mmol) in anv. DCM (33 ml) under argon atmosphere was added: HOAt (980 mg, 7.2 mmol), D-Leu-OBn.pTsOH (2.65 g, 12 mmol), NMM (1.21 g, 12 mmol) and DCC (1.48 g, 7.2 mmol). The mixture is stirred for 2 hours at 0 ° C and then 12 hours at rt .; additional D-Leu-OBn.pTsOH (0.66 g, 3 mmol) and NMM (0.30 g, 3 mmol) were added and the mixture was stirred for another 3 hours. The mixture was filtered and the solvent was concentrated in vacuo. The residue was dissolved in EtOAc (30 ml) and washed successively with NaHCO ₃ (2 × 25 ml, sat.), Citric acid (2 × 25 ml, 10%) and brine (25 ml). The organic solution was dried (Na ₂ SO ₄ ) and concentrated under reduced pressure. The residue was purified by LC-silica (hex. - EtOAc, 6: 1) to give 9 (2.63 g, 96%) as a colorless oil. [α] _D ²⁰ 12.4 ° (s 1, MeOH). HPLC [column Bondapack C ₁₈ (Waters), 10 m, 3.9 × 300 mm, flow: 1 ml / min, at 214 nm, CAN eluent / 0.05% TFA (40:60)] t _R = 9, 08 min].

¹ H-NMR (300 MHz, DMSO-d ₆ ) δ 0.86 (m, 6H), 1.37 (s, 9H), 1.55-1.72 (m, 5H), 2.00 (m , 2H), 2.64 (m, 1H), 2.86 (m, 1H), 3.15 (m, 1H), 3.55 (m, 1H), 4.41 (m, 1H), 5 05-5.11 (m, 4H); 5.63-5.72 (m, 1H); 7.28-7.35 (m, 5H).

¹³ C-NMR (75 MHz, acetone-d ₆ ) δ 21.6, 22.8, 24.6, 28.3, 34.7, 38.2, 41.3, 49.38, 51.0, 66.9, 69.7, 80.1, 119.1, 128.3, 132.7, 153.9, 172.8.

Link, synthesis 8: a) Seebach, D. et al. J. Am. Chem. Soc. 1983, 105, 5390-5398. b) Genin, M. J. et al. J. Org. Chem. 1993, 58, 2334-2337.

Example 85

Synthesis of (5R, 8RS) -1- (tert-butoxycarbonyl) -7 - [(1R) -1-benzyloxycarbonyl-3-methylbutyl] -8-hydroxy-6-oxo-1,7-diazaspiro [4.4] nonane (10 )

To a solution of 9 (1.56 g, 3.42 mmol) in MeOH / H ₂ O (2: 1, 108 ml) in an argon atmosphere, a solution of OsO ₄ (2.5% m / m, 2.9 ml) tert-butanol. Stirring was continued for another 10 minutes and NaIO ₄ (2.195 g, 10.3 mmol) was added. After 24 hours of stirring, the reaction mixture was diluted with H ₂ O (100 ml) and extracted with BtOAc (3 × 50 ml). The combined organic extracts were washed with brine (50 ml), dried (Na ₂ SO ₄ ) and concentrated under reduced pressure. The residue was purified by LC-silica (hex. EtOAc, gradient 80:20 - 0: 100%) to give diastereomers 10a and 10b (together: 1.17 g, 76%) as a white solid.

10a: HPLC [Column Novapack C ₁₈ (Waters), 3.9 × 150 mm, ⌀ = 1 ml / min, λ = 214 nm, eluent: CH ₃ CN / 0.05% TFA, (40/60)] t _R = 14.45 min. Mp: 140-141 ° C. [α] _D ²⁰ -4 ° (s 1, MeOH).

¹ H-NMR (300 MHz, acetone-d ₆ ) δ 0.90 (m, 6H), 1.29 (s, 9H), 1.64-2.30 (m, 9H); 2.68 [dd, 1H, J ₁ = 6, J ₂ = 13, 1H), 3.37 (m, 2H), 4.51 (dd, 1H), 5.13. (d, J = 15, 2H), 5.79 (t, J = 5, 2H), 7.40 (m, 5H).

¹³ C-NMR (75 MHz, acetone-d ₆ ) δ 21.8, 23.8, 24.1, 24.9, 28.5, 39.6, 40.8, 41.5, 48.5, 66.8, 79.4, 79.8, 81.2, 129.1, 129.3, 128.6, 171.7, 172.0.

ESI-MC Calculate. for C ₂₅ H ₃₆ N ₂ O ₆ : 460.26. Found m / z: 483.4 (M + Na) ⁺ .

10b: HPLC [Column Novapack C ₁₈ (Waters), 3.9 x 150 mm, ⌀ = 1 ml / min, λ = 214 nm, eluent: CH ₃ CN / 0.05% TFA, (40/60)] t _R = 18.75 min. Mp: 134-135 ° C. [α] _D ²⁰ +26 (c 1.2, MeOH).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.90 (6H, m), 1.40 (9H, s), 1.50-2.60 (9H, m), 3.40 (2H, m ), 4.20-5.40 (5H, m), 7.40 (5H, m).

¹³ C-NMR (75 MHz, acetone-d ₆ ) δ 21.3, 23.2, 24.1, 24.9, 28.3, 38.9, 40.2, 42.5, 47.9, 53.2, 66.8, 77.5, 79.4, 80.6, 129.1, 171.2, 174.1.

ESI-MC Calculate. for C ₂₅ H ₃₆ N ₂ O ₆ : 460.25. Found m / z: 483.5 (M + Na) ⁺ .

Example 86

Synthesis of (5R) -7 - [(1R) -1-benzyloxycarbonyl-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane as trifluoroacetate salt (11)

To a solution of 10 (430 mg, 0.93 mmol) in TFA (10 ml) was added NaBH ₄ (106 mg, 2.8 mmol). The mixture was stirred for 2 hours and then the reaction mixture was concentrated under reduced pressure. The residue was partitioned between H ₂ O (5 ml) and DCM (20 ml). The organic phase was dried (Na ₂ SO ₄ ) and concentrated in vacuo to give 11 as an orange oil (318 mg, quant.). [α] _D ²⁰ +15 (s 1, MeOH).

¹ H-NMR (300 MHz, acetone-d ₆ ) δ 0.91 (m, 6H), 1.50 (m, 1H), 1.66-1.94 (m, 2H), 2.11-2 72 (m, 6H), 3.48-3.72 (m, 4H), 4.74 (dd, J ₁ = 6, J ₂ = 15, 1H), 5.18 (s, 2H), 7 37 (m, 5H).

¹³ C-NMR (75 MHz, acetone-d ₆ ) δ 21.2, 23.2, 23.9, 25.5, 30.2, 34.6, 38.0, 42.2, 46.6, 53.9, 67.6, 69.6, 129.3, 161.1, 161.6, 170.9, 172.6.

ESI-MC Calculate. for C ₂₀ H ₂₈ N ₂ O ₃ : 344.21. Found m / z: 345.3 (M + H) ⁺ .

Example 87

Synthesis of (5R) -1- (tert-butoxycarbonyl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane (12)

To a solution of 11 (150 mg, 0.44 mmol) in ACN (5 ml) was added tetramethylammonium hydroxide pentahydrate (158 mg, 0.87 mmol) and Boc ₂ O (144 mg, 0.66 mmol) with stirring. After 6 hours, an additional amount of TMAN. 5 H ₂ O (158 mg) and Boc ₂ O (192 mg) were added. The reaction mixture was stirred for 2 hours and then partitioned between H ₂ O (10 ml) and DCM (25 ml). The aqueous phase is lyophilized and purified by LC-silica (DCM-MeOH, gradient 92: 8 - 60:40) to give 12 (100 mg, 64%) as a white solid.

Example 88

Synthesis of (5R) -1- (isobutyryl) -7 - [(1R) -1-benzyloxycarbonyl-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane (13)

To a solution of 11 (169 mg, 0.49 mmol) in anhydrous. DCM (10 ml) at 0 ° C. under argon atmosphere was added TEA (199 mg, 1.96 mmol), DMAP (6 mg, 0.049 mmol) and isobutyryl chloride (104 mg, 0.98 mmol) dropwise. The reaction mixture was allowed to warm to room temperature and stirred for 24 hours. The crude product was partitioned between H ₂ O (10 ml) and DCM (10 ml). The organic phase was washed with brine (10 ml), dried (Na ₂ SO ₄ ) and concentrated in vacuo. Pure compound 13 (150 mg, 74%) was obtained as a white solid after LC-silica (hex. EtOAc, 60:40 - 0: 100 gradient).

HPLC [Column Novapack C ₁₈ (Waters), 3.9 × 150 mm, ⌀ = 1 ml / min, λ = 214 nm, eluent: CH ₃ CN / 0.05% TFA, (50/50)] t _R = 4.50 minutes Mp: 87 ° C. [α] ²⁰ _D + 9.6 (s 1.4, MeOH).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.90 (2d, J = 7, 6H), 1.09-1.12 (2d, 6H), 1.37 (septet, J = 6, 1H) 1.61-2.10 (m, 7H), 2.64 (m, 2H), 3.14 (dd, J ₁ = 9, J ₂ = 17, 1H), 3.64 (m, 3H) 4.85 (dd, J ₁ = 5, J ₂ = 10, 1H), 5.14 (d, J = 6, 2H), 7.32 (m, 5H).

¹³ C-NMR (300 MHz, CDCl ₃ ) δ 18.6, 18.7, 21.2, 23.1, 24.0, 24.8, 29.5, 32.5, 35.7, 37, 6, 40.5, 47.8, 52.7, 66.7, 76.3, 170.8, 174.3, 174.9.

ESI-MC Calculate. for C ₂₁ H ₃₄ N ₂ O: 414.25. Found m / z: 415.4 (M + H) ⁺ .

Example 89

Synthesis of (5R) -1- (pyruvyl) -7 - [(1R) -1-benzyloxycarbonyl-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane (14)

Pyruvyl chloride is prepared according to the procedure described in the literature, Pansare, S.V .; Gnana R.R. "Asymmetric Allylation and reduction on an Ephedrine-Derived Template: Stereoselective Synthesis of α-Hydroxy Acids and Derivatives" J. Org. Chem. 1998, 63, 4120-4124. α, α-Dichloromethyl methyl ether (188 mg, 1.57 mmol) is added to pyruvic acid (115 mg, 1.31 mmol). The reaction mixture was stirred for 20 minutes and the resulting solution was heated to 50-55 ° C and then stirred for an additional 30 minutes. The reaction mixture was allowed to cool to room temperature and DCM (3 ml) was added.

To a solution of 11 (150 mg, 0.33 mmol) in bezv. DCM (4 ml) at 0 ° C in argon atmosphere add TEA (200 mg, 1.98 mmol), DMAP (4 mg, 0.033 mmol) and a freshly prepared solution of pyruvyl chloride at 0 ° C. The reaction mixture was allowed to warm to room temperature and was stirred for 6 hours. The crude product was washed successively with citric acid (5 ml, 10%), aq. NaHCO ₃ us. (5 ml) and brine (5 ml). The organic phase is dried (Na ₂ SO ₄ ) and concentrated. Pure compound 14 (77 mg, 56%), in the form of an oil, was obtained after LC-silica (hex. - EtOAc, 1: 3).

HPLC [Column Novapack C ₁₈ (Waters), 3.9 × 150 mm, ⌀ = 1 ml / min, λ = 214 nm, eluent: ACN / 0.05% TFA, (50/50)] t _R = 5, 87 and 6.72 minutes

¹ H-NMR (200 MHz, CDCl ₃ ) δ 0.92 and 0.95 (2d, J = 6.6H); 1.42 (m, 1H); 1.61-2.39 (m, 7H), 2.44 (s, 3H), 2.77 (m, 1H), 3.22 (m, 1H), 3.56-3.78 (m, 2H), 3.92 (m, 1H), 4.67 and 4.85 (dd, J ₁ = 6, J ₂ = 10, 1H), 5.21 (s, 2H), 7.34 (m, 5H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 21.2, 23.0, 24.4, 24.8, 26.4, 29.3, 35.6, 37.3, 40.7, 48, 9, 53.0, 66.8, 68.6, 135.0, 166.0, 170.8, 173.0, 198.0.

ESI-MC Calculate. for C ₂₃ H ₃₀ N ₂ O ₅ : 414.22. Found m / z: 415.4 (M + H) ⁺ .

Example 90

Synthesis of (5R) -1- (2-methylacryloyl) -7 - [(1R) -1-benzyloxycarbonyl-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane (15)

Following the procedure developed for synthesis 13, starting from 11 (200 mg, 0.43 mmol) and methyl acryloyl chloride (89 mg, 0.86 mmol), the title compound (70 mg, 50%) was obtained as a colorless oil, after purification LC (silica gel, hex. - EtOAc, 2: 1). HPLC [Column Novapack C18 (Waters), 3.9 × 150 mm, ⌀ = 1 ml / min, λ = 214 nm, eluent: ACN / 0.05% TFA, (25/75)]. Rt = 6.38 min.

¹ H-NMR (200 MHz, CDCl ₃ ) δ 0.91 (t, J = 6, 6H), 1.44 (m, 1H), 1.64-1.93 (m, 5H), 1.93 (s, 3H), 1.96-2.12 (m, 2H), 2.78 (m, 1H), 3.20 (m, 1H), 3.56-3.68 (m, 3H), 4.80 and 4.77 (2d, J = 10, 1H), 5.16 (s, 2H), 5.19 (d, J = 9, 2H), 7.32 (m, 5H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 19.8, 21.4, 23.0, 24.2, 24.9, 29.9, 37.1, 37.7, 41.1, 50, 2, 53.2, 66.6, 67.4, 116.9, 128.1, 128.3, 128.4, 135.0, 141.7, 170.7, 174.1.

ESI-MC Calculate. for C ₂₄ H ₃₂ N ₂ O ₄ : 412.24. Found m / z: 413.3 (M + H) ⁺ .

Example 91

Synthesis of (5R) -1- (isobutyryl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane (16)

A degassed solution of 13 (134 mg, 0.32 mmol) in methanol containing 10% Pd / C (27 mg) was hydrogenated at 16 psi for 24 hours. The mixture was filtered through a pad of celite and the filtered solution was concentrated under reduced pressure. receiving 16 (100 mg, 95%) as a colorless oil. Mp 68-69 ° C. [α] ²⁰ _D -2 ° (c 1.1, MeOH).

¹ H-NMR (300 MHz, acetone-d ₆ ) δ 0.87-0.91 (2d, J = 7.6H), 1.09-1.12 (2d, 6H), 1.46 (m, 1H), 1.70 (m, 2H), 1.90-2.10 (m, 5H), 2.49 (m, 1H), 2.70 (m, 1H), 3.30 (m, 1H ), 3.49 (dt, J ₁ = 8, J ₂ = 10, 1H), 3.61-3.75 (m, 2H), 4.71 (dd, J ₁ = 5, J ₂ = 11, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 16.4, 16.6, 21.1, 23.0, 24.3, 25.0, 30.8, 32.6, 36.6, 36, 9, 40.6, 48.0, 53.4, 67.5, 171.8, 174.2, 176.2.

ESI-MC Calculate. for C ₁₇ H ₂₈ N ₂ O ₄ : 324.20. Found: 323.3 (M-1) ⁺ .

Example 92

Synthesis of (5R) -1- (pyruvyl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane (17)

A degassed solution of 14 (79 mg, 0.26 mmol) in methanol (20 ml) containing Pd / C (10%, 22 mg) was hydrogenated at atmospheric pressure for 45 minutes. The filtered solution was concentrated under reduced pressure to obtain 17 (79 mg, 95%) as a white solid. HPLC [Column Novapack C ₁₈ (Waters), 3.9 × 150 mm, ⌀ = 1 ml / min, λ = 214 nm, eluent: ACN / 0, 05% TFA, (20/80)]. Rt = 13.14 min.

¹ H-NMR (200 MHz, CDCl ₃ ) δ 0.93 (m, 6H), 1.43 (m, 1H), 1.71-2.22 (m, 7H), 2.34 (s, 3H ), 2.41 (s, 3H), 2.74 (m, 1H), 3.31 (m, 1H), 3.74 (m, 2H), 3.92 (m, 1H), 4.80 (dd, J ₁ = 6, J ₂ = 10, 1H), 7.07 (br s, 1H).

¹³ C-NMR (200 MHz, CDCl ₃ ) δ 21.3, 23.2, 24.7, 25.1, 27.1, 29.9, 36.0, 37.0, 41.0, 49, 3, 53.6, 69.1, 162.1, 172.7, 173.7, 197.5.

ESI-MC Calculate. for C ₂₃ H ₃₀ N ₂ O ₅ : 324.17. Found m / z: 325.1 (M + H) ⁺ .

Example 93

Synthesis of (5R) -1- (2-methylacryloyl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane (18)

To a solution of 15 (65 mg, 0.16 mmol) in methanol (2.5 ml) was added aq. NaOH (1.6 ml, 1 N) and H ₂ O (1.6 ml) and the mixture was stirred at room temperature for 1 hour. The resulting mixture was concentrated under reduced pressure and the residue was partitioned between H ₂ O (20 ml) and DCM (20 ml). The aqueous phase is acidified to pH 2 with aq. HCl (10 ml, 0.1 N) and extracted with DCM (3 × 20 ml). The combined organic phases were washed with brine (25 ml), dried (Na ₂ SO ₄ ) and concentrated under reduced pressure to obtain 18 (44 mg, 85%) as a white solid. HPLC [Column Novapack C ₁₈ (Waters), 3.9 × 150 mm, ⌀ = 1 ml / min, λ = 214 nm, eluent: ACN / 0.05% TFA, (25/75)] R _t = 6, 38 minutes

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.86 (d, J = 5, 3H), 0.89 (d, J = 5, 3H), 1.42 (m, 1H), 1.55 -2.25 (m, 8H), 2.56 (m, 1H), 3.19-3.44 (m, 2H), 3.57-3.67 (m, 2H), 4.78 (d , J = 11, 1H), 4.82 (d, J = 11, 1H), 5.18 (d, J = 6, 2H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 19.5, 21.2, 23.1, 24.2, 25.1, 30.9, 36.5, 37.6, 40.9, 50, 5, 53.7, 67.5, 117.5, 140.6, 170.8, 174.2.

ESI-MC Calculate. for C ₁₇ H ₂₆ N ₂ O ₄ : 322.19. Found: 323.2 (M + H) ⁺ .

Example 94

Synthesis of [(5R) -1- (tert-butoxycarbonyl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane ^7-9 aplidine (9SBSAPL1)

Following the procedure developed for the synthesis of SAPL3, starting from SAPL4 (10 mg, 13 μmol), 12 (5 mg, 14 μmol), HATU (12.4 mg), HOAt (4.5 mg), NMM (3.3 μl ), DCM (140 μl) and DMF (70 μl), the title compound (11 mg, 73%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, ACN / H ₂ O ratio 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, R _t = 30 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.85-0.97 (m, 24H), 1.19-1.34 (m, 18H), 1.48 (s, 9H), 1.50 -2.20 (m, 6H), 2.32-2.36 (m, 1H), 2.54 (s, 3H), 2.58-2.72 (m, 2H), 2.97-3 08 (m, 1H), 3.10-3.22 (m, 3H), 3.34 (dd, 1H, J = 3.9, 13.8), 3.46-3.79 (m, 6H), 3.79 (s, 3H), 4.03-4.12 (m, 2H), 4.28 (dd, 1H, J = 6.6, 13.2), 4.57-4, 63 (m, 2H), 4.79-4.88 (m, 2H), 5.15 (d, 1H, J = 3.3), 5.21-5.23 (m, 1H), 6, 84 (d, 2H, J = 8.4), 7.07 (d, 2H, J = 8.7), 7.28 (d, 1H, J = 10.8), 7.79 (d, 1H J = 6.6), 7.82 (d, 1H, J = 9.9).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.66, 15.21, 15.43, 16.79, 17.23, 18.69, 18.77, 21.24, 23.67, 23, 95, 24.10, 24.90, 25.10, 25.38, 27.04, 28.23, 28.75, 29.93, 31.61, 34.50, 33.88, 34.16, 36.20, 36.57, 38.76, 39.09, 39.78, 41.29, 47.27, 47.76, 49.60, 49.96, 52.66, 55.50, 56, 26, 57.38, 58.18, 66.74, 66.86, 68.34, 70.53, 80.75, 81.93, 114.33, 130.23, 130.54, 168.08, 169.83, 170.18, 170.80, 171.43, 172.55, 175.04, 205.04.

ESI-MC Calculate. for C ₆₀ H ₉₃ N ₇ O ₁₅ 1151.7. Found m / z: 1152.4 (M + H) ⁺ .

Example 95

Synthesis of [Hiv] ³ - [(5R) -1- (tert-butoxycarbonyl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane] ^{7 -9-} aplidine (9SBSHPL1)

Following the procedure developed for the synthesis of SAPL3, starting from SHPL4 (10 mg, 13 μmol), 12 (5 mg, 14 μmol), HATU (14 mg), HOAt (5 mg), NMM (6 μl), DCM (150 μl ) and DMF (50 μl), the title compound (10 mg, 70%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, iso-ratio ACN / H ₂ O 85:15 (flow: 7 ml / min, 250 × 21 mm, at 270 nm, R _t = 28.1 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.80-1.07 (m, 24H), 1.08-1-67 (m, 12H), 1.48 (s, 9H), 1.68 -2.30 (m, UN), 2.41 (m, 1H), 2.55 (s, 3H), 2.68 (m, 1H), 2.94 (m, 1H), 3.07- 3.40 (m, 4H), 3.42-3.72 (m, 6H), 3.78 (s, 3H), 3.90 (m, 1H), 4.01 (m, 1H), 4 29 (m, 1H), 4.63 (m, 1H), 4.77 (m, 1H), 4.87 (m, 1H), 5.02 (d, J = 4.8, 1H), 5.25 (m, 1H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8.3, 2H), 7.31 (d, J = 9.7 , 1H), 7.50 (d, J = 5.8, 1H), 7.85 (d, J = 9.7, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 12.07, 14.25, 17.12, 17.92, 19.15, 21.11, 21.18, 23.80, 24.00, 24, 06, 24.86, 25.08, 25.15, 27.58, 28.21, 28.82, 30.33, 31.18, 33.78, 34.28, 35.77, 36.65, 39.07, 39.45, 39.86, 46.91, 48.14, 48.47, 52.62, 55.49, 57.13, 58.42, 66.36, 66.80, 69, 14, 70.84, 79.19, 80.55, 114.27, 130.29, 130.59, 154.12, 158.81, 168.25, 169.84, 170.72, 170.80, 170.90, 171.25, 174.89.

ESI-MC Calculate. for C ₅₇ H ₈₉ N ₇ O ₁₄ 1095.6. Found m / z: 1096.9 (M + H) ⁺ .

Example 96

Synthesis of [(5R) -1- (isobutyryl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane] ^7-9 aplidine (9SISAPL1)

Following the procedure developed for the synthesis of SAPL3, starting from SAPL4 (11 mg, 12.9 μmol), 16 (5 mg, 15.4 μmol), HATU (14 mg), HOAt (5 mg), NMM (3.6 μl ), DCM (155 μl) and DMF (78 μl), the title compound (10 mg, 69%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, ACN / H ₂ O ratio 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 19 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.86-1.00 (m, 24H), 1.12 (d, J = 6.9, 3H), 1.18 (d, J = 6, 6, 3H), 1.34 (t, J = 6.6, 2H), 0.90-1.30 (m, 7H), 1.56-2.25 (m, 16H), 2.30- 2.80 (m, 3H), 2.55 (s, 3H), 2.95-3.06 (m, 1H), 3.15-3.25 (m, 3H), 3.65 (dd, 1H), 3.52-3.79 (m, 6H), 3.79 (s, 3H), 3.98-4.15 (m, 1H), 4.28 (dd, J ₁ = 6.6 , J ₂ = 10.3, 1H), 4.59 (m, 2H), 4.79-4.85 (m, 2H), 5.17 (d, J = 3.6, 1H), 5, 40-5.44 (m, 1H), 6.84 (d, J = 8.4, 2H), 7.06 (d, J = 8.7, 2H), 7.24 (d, J = 11 , 1, 1H), 7.90 (d, J = 9.3, 1H), 8.56 (d, J = 5.1, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.50, 14.92, 15.22, 16.68, 16.95, 18.50, 18.59, 18.81, 20.94, 23, 42, 23.80, 24.49, 24.68, 24.90, 25.11, 26.91, 27.98, 30.93, 31.30, 35.58, 33.88, 34.16 35.85, 36.24, 38.64, 38.84, 39.71, 41.29, 47.01, 47.76, 49.42, 49.62, 52.66, 55.26, 55, 73, 57.12, 58.21, 66.57, 67.40, 68.10, 70.79, 81.57, 114.07, 130.05, 130.31, 158.57, 168.12, 169.66, 170.08, 170.56, 171.13, 171.96, 172.37, 174.04, 175.41, 205.04.

ESI-MC Calculate. for C ₅₉ H ₉₁ N ₇ O ₁₄ : 1121.66. Found: 1122.8 (M + H) ⁺ .

Example 97

Synthesis of [Hiv] ³ - [(5R) -1- (tert-butoxycarbonyl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane] ^{7 -9-} aplidine (9SISHPL1)

Following the procedure developed for the synthesis of SAPL3, starting from SHPL4 (10 mg, 13 μmol), 16 (4.5 mg, 14 μmol), HATU (14 mg), HOAt (5 mg), NMM (6 μl), DCM ( 150 μl) and DMF (50 μl), the title compound (10 mg, 72%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, iso-ratio of ACN / H ₂ O 85:15 (flow: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 16.9 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.80-1.07 (m, 24H), 1.08-1.47 (m, 12H), 1.48-2.30 (m, 16H) 2.36 (m, 2H), 2.56 (s, 3H), 2.65 (m, 1H), 2.96 (m, 1H), 3.18 (m, 2H), 3.36 ( m, 2H), 3.65 (m, 6H), 3.78 (s, 3H), 3.91 (m, 1H), 4.02 (m, 1H), 4.25 (m, 1H) 4 63 (m, 1H), 4.72 (m, 2H), 4.87 (m, 1H), 5.02 (d, J = 4.8, 1H), 5.45 (m, 1H), 6.84 (d, J = 8.7, 2H), 7.07 (d, J = 8.7 2H), 7.27 (d, J = 4.8, 1H), 7.88 (d, J = 9.7, 1H); 8.32 (d, J = 4.8, 1H).

ESI-MC Calculate. for C ₅₆ H ₈₇ N ₇ O ₁₃ 1065.6. Found m / z: 1066.7 (M + H) ⁺ .

Example 98

Synthesis of [(5R) -1- (pyruvyl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane] ^7-9 aplidine (9SPSAPL1)

Following the procedure developed for the synthesis of SAPL3, starting from SAPL4 (10 mg, 11.7 μmol), 17 (5 mg, 15.4 μmol), HATU (12 mg), HOAt (5 mg), NMM (5 μl), DCM (140 μl) and DMF (70 μl), the title compound (9 mg, 63%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, iso-ratio ACN / H ₂ O 85:15 (flow rate : 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 14.4 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.90-1.00 (m, 24H), 1.05-1.40 (m, 12H), 1.40-2.25 (m, 16H) 2.27-2.41 (m, 1H), 2.42-2.70 (m, 3H), 2.54 (s, 3H), 2.92-2.98 (m, 1H), 3 , 12-3-38 (m, 4H), 3.54-3.78 (m, 4H), 3.79 (s, 3H), 4.01-4.12 (m, 2H), 4.20 -4.26 (m, 2H), 4.57-4.62 (m, 2H), 4.77-4.82 (m, 2H), 535.18 (d, J = 3.0, 2H) 5.37-5.42 (m, 1H), 6.84 (d, J = 8.7, 2H), 7.07 (d, J = 8.7, 2H), 7.20 (d, J = 9.6, 1H), 7.85 (d, J = 9.6, 1H), 98.04 (d, J = 5.4, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.86, 14.97, 15.47, 16.78, 17.12, 18.84, 21.20, 21.27, 23.62, 24, 14, 15.03, 25.15, 25.30, 27.35, 27.51, 28.19, 30.46, 31.52, 34.27, 35.95, 39.01, 40.07, 41.59, 47.25, 49.72, 53.08, 55.51, 55.81, 57.38, 58.21, 66.66, 68.19, 69.23, 70.74, 81, 69, 85.15, 114.33, 130.13, 130.56, 158.85, 161.12, 168.49, 169.81, 169.91, 170.81, 171.38, 171.69, 172.60, 173.34, 197.64, 205.19.

ESI-MC Calculate. for C ₅₈ H ₈₇ N ₇ O ₁₅ : 1121.63. Found 1122.3 (M + H) ⁺ .

Example 99

Synthesis of [Hiv] ³ - [(5R) -1- (pyruvyl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane] ^7-9 aplidine (9SPSHPL1)

Following the procedure developed for the synthesis of SAPL3, starting from SHPL4 (10 mg, 13 μmol), 17 (4.5 mg, 14 μmol), HATU (14 mg), HOAt (5 mg), NMM (6 μl), DCM ( 150 μl) and DMF (50 μl), the title compound (10 mg, 72%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, iso-ratio of ACN / H ₂ O 85:15 (flow: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 13.6 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.80-1.10 (24H, m), 1.11-1.80 (12H, m), 1.81-2.30 (10H, m) 2.45 (m, 1H), 2.55 (s, 3H), 2.57 (s, 3H), 3.07-3.43 (m, 6H), 3.52-3.77 (m 6H), 3.78 (s, 3H), 3.91 (m, 1H), 4.03 (m, 1H), 4.29 (m, 1H), 4.63 (m, 1H), 4 72 (m, 1H), 4.87 (m, 1H), 5.03 (d, J = 4.3, 1H), 5.45 (m, 1H), 6.84 (d, J = 8 , 3, 2H), 7.07 (d, J = 8.3, 2H), 7.29 (d, J = 8.7, 1H), 7.81 (d, J ₁ = 9.2, 1H ), 7.87 (d, J = 4.8, 1H).

ESI-MC Calculate. for C ₅₅ H ₈₃ N ₇ O ₁₄ : 1065.6. Found 1066.4 (M + H) ⁺ .

Example 100

Synthesis of [Hiv] ³ - [(5R) -1- (acryloyl) -7 - [(1R) -1-carboxy-3-methylbutyl] -6-oxo-1,7-diazaspiro [4.4] nonane] ^7-9 aplidine (9SASHPL1)

Following the procedure developed for the synthesis of SAPL3, starting from SHPL4 (10 mg, 13 μmol), 18 (5.8 mg, 14 μmol), HATU (14 mg), HOAt (5 mg), NMM (6 μl), DCM ( 150 μl) and DMF (50 μl), the title compound (10 mg, 72%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, iso-ratio of ACN / H ₂ O 85:15 (flow: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 16.4 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.85-0.96 (m, 18H), 1.02-1.05 (m, 6H), 1.14-1.45 (m, 12H) 1.49-1.64 (m, 4H), 1.68-1.77 (m, 1H), 1.89-2.05 (m, 3H), 1.99 (s, 3H), 2 10-2.28 (m, 4H), 2.43 (dd, J ₁ = 7.8, J ₂ = 17.1, 1H), 2.57 (s, 3H), 2.60-2, 68 (m, 1H), 2.97 (br.s, 1H), 3.13-3.40 (m, 4H), 3.54-3.77 (m, 5H), 3.79 (s, 3H), 3.89-4.07 (m, 2H), 4.27 (m, 1H), 4.64 (m, 1H), 4.73 (m, 1H), 4.88 (m, 1H ), 5.03 (d, J = 4.4, 1H), 5.30 (d, J = 20, 1H), 5.30-5.39 (m, 1H), 6.84 (d, J = 8.3, 2H), 7.08 (d, J = 8.3, 2H), 7.29 (s, 1H), 7.88 (d, J = 9.8, 1H), 8.23 (d, J = 7.4, 1H).

ESI-MC Calculate. for C ₅₆ H ₈₅ N ₇ O ₁₃ : 1063.6. Found 1064.6 (M + H) ⁺ .

Example 101

Synthesis of [Hiv] ³ - [Z-Ala] ⁹ -aplidine (9ZASHPL2)

To a flask containing Z-Ala-Pro-OH (36 mg, 112 μmol) in DCM (0.4 ml) at 0 ° C under argon, DIPCDI (10 μl, 64 μmol) was added and the mixture was stirred for 60 minutes. Then, a solution of SHPL2 (20 mg, 22.5 μmol) in DCM (0.2 ml) was added and after 3 hours the reaction was quenched by addition of aq. HCl (3 ml, 0.1 N). The mixture was stirred for 5 minutes and then diluted with DCM (4 ml) and washed successively with aq. KHSO ₄ (4 ml, 10%), aq. NaHCO ₃ (4 ml, sat.) And brine (4 ml). The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. Purification of the residue by HPLC (HyperPrep PEP 100 C18, isotrate ratio ACN / H ₂ O 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 18.2 min) gives 9ZASHPL2 (30 mg, 67%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 24H), 1.12-1.50 (m, 18H), 1.52-2.70 (m, 6H) 2.45 (m, 1H), 2.56 (s, 3H), 2.96-3.38 (m, 4H), 3.10 (s, 3H), 3.52-3.72 (m 5H), 3.85 (m, 1H), 3.78 (s, 3H), 4.01 (m, 1H), 4.18 (m, 1H), 4.51 (m, 1H), 4 64 (m, 1H), 4.71 (m, 1H), 4.87 (m, 1H), 5.02 (d, J ₁ = 5.3, 1H), 5.06 (m, 2H) 5.25 (m, 1H), 5.42 (m, 1H), 6.10 (d, J = 8.3, 1H), 6.84 (d, J = 8.3, 2H), 7 07 (d, J = 8.3, 2H), 7.31 (m, 6H), 7.70 (d, J = 4.3, 1H), 7.77 (d, J = 9.7, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.80, 13.97, 16.63, 17.02, 17.48, 18.93, 20.86, 21.15, 23.45, 23, 76, 24.78, 25.96, 27.31, 27.92, 28.50, 30.06, 31.36, 33.44, 33.82, 35.66, 38.70, 39.38, 39.63, 46.54, 46.62, 46.98, 47.10, 48.19, 48.59, 54.89, 54.97, 55.22, 56.44, 56.90, 58, 14, 66.16, 66.36, 68.95, 70.94, 78.78, 114.00, 127.75, 128.30, 129.90, 130.26, 156.31, 158.53, 168.67, 169.57, 170.06, 170.25, 170.65, 171.97, 173.08, 174.67.

ESI-MC Calculate. for C ₆₂ H ₉₂ N ₈ O ₁₅ 1188.67. Found m / z 1189.7 (M + H) ⁺ .

Example 102

Synthesis of [Hiv] ³ - [Vos-Ala] ⁹ -aplidine (9BASHPL2)

Following the procedure developed for the synthesis of 9ZASHPL2, from SHPL2 (10 mg, 11.2 μmol), Boc-Ala-Pro-OH (17 mg, 57 μmol), DIPCDI (5 μl) and DCM (300 μl), indicated in the title the compound (9 mg, 70%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, iso ratio ACN / H ₂ O 85:15 - 100: 1 in 10 min (flow: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 14.5 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 24H), 1.12-1.50 (m, 15H), 1.40 (s, 9H), 1.52 -2.70 (m, 9H), 2.45 (m, 1H), 2.60 (s, 3H), 3.00-3.43 (m, 4H), 3.10 (s, 3H), 3.62 (m, 5H), 3.79 (s, 3H), 3.90 (m, 1H), 4.02 (m, 1H), 4.20 (m, 1H), 4.41 (m , 1H), 4.67 (m, 2H), 4.87 (m, 1H), 5.02 (d, J ₁ = 4.3, 1H), 5.26 (m, 1H), 5.40 (m, 1H), 5.76 (d, J = 7.8, 1H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8.3, 2H) 7.32 (d, J = 9.7, 1H); 7.73 (d, J = 4.8, 1H); 7.80 (d, J = 9.7, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 12.06, 14.34, 16.88, 17.20, 27.77, 19.17, 21.10, 21.41, 23.72, 24, 03, 25.06, 26.17, 27.60, 28.19, 28.60, 30.31, 31.63, 33.74, 35.96, 38.96, 38.93, 39.59, 39.86, 46.86, 47.27, 48.29, 48.45, 55.18, 55.42, 55.49, 56.59, 57.16, 58.44, 66.43, 69, 14, 71.30, 79.05, 79.40, 114.29, 130.21, 130.54, 156.06, 158.83, 168.80, 169.87, 170.51, 170.64, 170.94, 171.31, 172.59, 173.45, 174.92.

ESI-MC Calculate. C ₅₉ H ₉₄ N ₈ O ₁₅ 1154.68. Found m / z 1155.6 (M + H) ⁺ .

Example 103

Synthesis of [Hiv] ³ - [Ala] ⁹ -aplidine (9ASHPL1)

In a flask containing 9BASHPL2 (8 mg, 6.92 μmol), add a solution of hydrochloric acid in bezv. dioxane (1.5 ml, 5.3 n, 7.9 mmol). The resulting solution was stirred at room temperature for 5 hours or until the starting material (TLC) completely disappeared. Then, the solution was concentrated under reduced pressure, and the residue was dissolved in DCM and concentrated again. White foamy crude product precipitated with DCM / hex. (2 ml / 4 ml) to give 9ASHPL1 (7.2 mg, quant.) As a white solid.

ESI-MC Calculate. for C ₅₄ H ₈₆ N ₈ O ₁₃ 1054.6. Found m / z: 1055.6 (M + H) ⁺ .

Example 104

Synthesis of [Hiv] ³ - [Boc-Pro] ^8- didemnin A (8PSHPL2)

Following the procedure developed for the synthesis of 9ZASHPL2, starting from SHPL2 (10 mg, 11.2 μmol), Boc-Pro-OH (13 mg, 57 μmol), DIPCDI (5 μl) and DCM (300 μl), the title compound (9 mg, 74%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, gradient ACN / H ₂ O 85:15 - 100: 0 in 10 min (flow rate: 7 ml / min, 250 × 21 mm , at 270 nm, t _R = 18.7 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 24H), 1.12-2.30 (m, 22H), 1.47 (s, 9H), 2.41 (m, 1H), 3.04 (s, 3H), 3.10-3.74 (m, 7H), 3.78 (s, 3H), 3.91 (m, 1H), 4.01 ( m, 1H), 4.31 (m, 1H), 4.59 (m, 1H), 4.87 (m, 1H), 5.02 (d, J = 4.8, 1H), 5.16 (m, 1H), 5.36 (m, 1H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8.3, 2H), 7.17 (d J = 6.3, 1H); 7.35 (d, J = 9.7; 1H); 7.85 (d, J = 9.7; 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 12.04, 14.41, 16.76, 17.97, 19.12, 21.03, 21.66, 23.92, 24.00, 24, 77, 25.09, 25.20, 27.53, 28.21, 28.72, 29.67, 30.35, 31.23, 33.80, 34.30, 36.24, 39.05, 39.37, 39.80, 46.90, 47.33, 48.42, 54.48, 55.49, 55.58, 55.84, 57.12, 58.08, 66.31, 69, 11, 71.05, 79.23, 80.28, 114.29, 130.23, 130.59, 154.91, 158.82, 168.37, 169.88, 170.66, 170.86, 171.28, 171.40, 174.10, 174.79.

ESI-MC Calculate. for C ₅₆ H ₉₁ N ₇ O ₁₄ : 1085.6. Found m / z: 1086.7 (M + H) ⁺ .

Example 105

Synthesis of [Hiv] ³ - [Pro] ^8- didemnin A (8PSHPL1)

Following the procedure developed for the synthesis of 9ASHPL1, starting from 8PSHPL2 (7 mg, 6.4 μmol), the title compound (6 mg, quant.) Was obtained as a white solid.

ESI-MC Calculate. for C ₅₁ H ₈₁ N ₇ O ₁₂ : 983.59. Found m / z: 984.6 (M + H) ⁺ .

Example 106

Synthesis of [Hiv] ³ - [Boc-Val] ^8- didemnin A (8VSHPL2)

Following the procedure developed for the synthesis of 9ZASHPL2, starting from SHPL2 (10 mg, 11.2 μmol), Boc-Val-OH (12 mg, 56 μmol), DIPCDI (5 μl) and DCM (300 μl), the title compound (9 mg, 82%) was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, gradient ACN / H ₂ O 85:15 - 100: 0 in 10 min (flow rate: 7 ml / min, 250 × 21 mm , at 270 nm, t _R = 22.1 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 30H), 1.12-1.82 (m, 12H), 1.45 (s, 9H), 1.84 -2.40 (m, 6H), 2.56 (s, 3H), 2.96 (s, 3H), 2.97 (m, 3H), 3.13 (m, 1H), 3.35 ( m, 1H), 3.56 (m, 1H), 3.65 (m, 2H), 3.79 (s, 3H), 3.85 (m, 1H), 4.03 (m, 1H), 4.24 (m, 1H), 4.41 (m, 1H), 4.61 (m, 1H), 4.88 (m, 1H), 4.99 (d, J ₁ = 5.3, 1H) ), 5.07 (m, 1H), 5.19 (m, 1H), 5.60 (d, J = 8.3, 1H), 6.84 (d, J = 8.3, 2H), 7.03 (d, J = 8.7, 2H), 7.07 (d, J = 8.3, 2H), 7.46 (d, J = 10.2, 1H), 7.84 (d , J = 9.2, 1H).

ESI-MC Calculate. for C ₅₆ H ₉₁ N ₇ O ₁₄ : 1085.6. Found m / z: 1086.7 (M + H) ⁺ .

Example 107

Synthesis of [Hiv] ³ - [Val] ^8- didemnin A (8VSHPL1)

Following the procedure developed for the synthesis of 9ASHPL1, starting from 8VSHPL2 (8 mg, 7.4 μmol), the title compound (7 mg, quant.) Was obtained as a white solid.

ESI-MC Calculate. for C ₅₁ H ₈₃ N ₇ O ₁₂ : 985.61. Found (m / z): 986.6 (M + H) ⁺ .

Example 108

Synthesis of [Hiv] ³ - [Val] ⁸ - [Isobutyryl] ⁹ Didemnin A (8V9ISHPL1)

To a solution of 8VSHPL1 (6 mg, 5.8 μmol) in DCM (200 μl) at 0 ° C in an Ar atmosphere was added NMM (3.3 μl, 30 μmol) and isobutyryl chloride (2 μl, 19 μmol). After 5 hours stirring at rt the reaction mixture was diluted with DCM (5 ml) and washed successively with water. KHSO ₄ (5 ml, 10%), aq. HCO ₃ Na (5 ml, sat.) And brine (5 ml). The organic solution was dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to obtain 8V9ISHPL1 (6 mg, 97%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.82-1.10 (m, 36H), 1.12-1.85 (m, UN), 1.90-2.45 (m, 6H) 2.56 (s, 3H), 2.90 (m, 1H), 3.00 (s, 3H,), 3.13 (m, 2H), 3.36 (dd, J ₁ = 4.3 , J ₂ = 14.1, 1H), 3.64 (m, 6H), 3.78 (s, 3H), 3.87 (m, 1H), 3.97 (m, 1H), 4.21 (m, 1H), 4.34 (m, 2H), 4.61 (m, 1H), 4.87 (m, 1H), 4.98 (d, J = 4.8, 1H), 5, 17 (m, 2H), 6.24 (d, J = 6.3, 1H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8.3, 2H ), 7.32 (d, J = 6.3, 1H), 7.35 (d, J = 4.3, 1H), 7.81 (d, J = 9.7, 1H).

ESI-MC Calculate. for C ₅₅ H ₈₉ N ₇ O ₁₃ , 1055.6. Found m / z: 1056.7 (M + H) ⁺ .

Example 109

Synthesis of [Coumarin] ⁸ Didemnin A (8CSAPL1)

Following the procedure developed for the synthesis of 9ZASHPL2, starting from SAPL2 (20 mg, 21 μmol) and coumarin-3-carboxylic acid (20 mg, 107 μmol), the title compound (18 mg, was obtained as a white solid after purification by HPLC (HyperPrep PEP 100 C18, isotrate ratio ACN / H ₂ O 85:15 (flow rate: 7 ml / min, 250 × 21 mm, at 270 nm, t _R = 16.5 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.78-1.00 (m, 24H), 1.20-2.50 (m, 22H), 2.56 (s, 3H), 2.92 (s, 3H), 3.08-3.25 (m, 2H), 3.90 (m, 1H), 3.60 (m, 2H), 3.70 (m, 1H), 3.79 ( s, 3H), 3.92-4.25 (m, 3H), 4.60 (m, 1H), 4.80 (m, 2H), 5.15 (m, 1H), 5.18 (d , J = 3.4, 1H), 5.28 (m, 1H), 6.84 (d, J = 8.3, 2H), 7.08 (d, J = 8.3, 2H), 7 20 (d, J = 6.3, 1H), 7.37 (m, 2H), 7.45 (d, J = 9.7, 1H), 7.58 (m, 2H), 7.96 (m, 1H); 8.23 (m, 1H).

ESI-MC Calculate. for C ₅₉ H ₈₂ N ₆ O ₁₅ : 1114.58. Found: 1116.3 (M + H) ⁺ .

Example 110

Synthesis of Coumarin-3-Carbonylaminoacetic Acid Methyl Ester (8G9C2)

Into a round bottom flask containing methylglycine (89 mg, 1.00 mmol), 3-carboxycoumarin, bezv. DCM (25 ml), in an Ar atmosphere, was added at room temperature N '- (3-dimethylaminopropyl) -N-ethylcarbodiimide hydrochloride (EDC) (479 mg, 2.50 mmol) and DMAP (489 mg, 4.00 mmol). The resulting mixture was stirred for 1 h 30 min (complete conversion by TLC). Then, DCM (20 ml) was added and the solution was washed successively with aq. NaHCO ₃ (10 ml, sat.) And brine (10 ml). The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated. The resulting orange solid was purified by flash LC (silica gel, hex.: EtOAc 1: 1 - 2: 1), which afforded the title compound (445 mg, quant.) As a colorless oil. Rf = 0.08 (hex. / BtOAc 1: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 3.79 (s, 3H), 4.25 (d, J = 5.4, 2H), 7.40 (m, 2H), 7.74 (d J = 7.3, 1H); 7.68 (m, 2H); 8.91 (s, 1H); 9.25 (s, 1H).

ESI-MC Calculate. for C ₁₃ H ₁₁ NO ₅ : 261.06. Found: 283.1 (M + Na) ⁺ .

Example 111

Synthesis of Coumarin-3-Carbonylaminoacetic Acid (8G9C1)

To a solution of coumarin-3-carbonylaminoacetic acid methyl ester (312 mg, 1.19 mmol) in THF (12 ml) in an Ar atmosphere at 0 ° C (ice bath) was added dropwise a solution of LiOH in H ₂ O (0.2 M ) The reaction mixture was vigorously stirred at room temperature until complete conversion by TLC (2 hours). The solution was partially concentrated and Et ₂ O (10 ml) was added. The organic layer was washed with NaHCO ₃ (10 ml, sat.) And the combined aqueous layers were acidified with 10% KHSO ₄ (pH 3-4) and extracted with diethyl ether (3 × 20 ml). The organic layer was concentrated under reduced pressure to obtain 8G9C1 (280 mg, 95%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 4.18 (d, J = 5.3, 2H), 7.41 (m, 1H), 7.45 (d, J = 7.8, 1H) 7.74 (d, J = 7.3, 1H), 7.78 (t, J = 8.3, 1H), 7.85 (d, J = 7.8, 1H], 8.89 ( s, 1H); 9.41 (m, 1H).

ESI-MC Calculate. for C ₁₂ H ₉ NO ₅ : 247.05. Found: 248.0 (M + H) ⁺ .

Example 112

Synthesis of [Gly] ⁸ - [Coumarin] ⁹ Didemnin A (8G9CSAPL1)

Following the procedure developed for the synthesis of 9ZASHPL2, starting from SAPL2 (20 mg, 21 μmol) and coumarin-3-carboxylic acid (26 mg, 105 μmol), the title compound (18 mg, 72%) was obtained as a white solid after purification by HPLC (Symetry Prep ™ C18, iso-ratio of ACN / H ₂ O of 60:40 (flow rate: 3 ml / min, 150 × 7.8 mm, at 270 nm, t _R = 17.5 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.85-0.94 (m, 24H), 1.23-2.17 (m, 21H), 2.30-2.44 (m, 1H) 2.56 (s, 3H), 3.01 (s, 3H), 3.10-3.24 (m, 2H), 3.40 (dd, J ₁ = 5.4, J ₂ = 14, 2, 1H), 3.59-3.74 (m, 3H), 3.79 (s, 3H), 3.99-4.21 (m, 4H), 4.37-4.44 (m, 1H), 4.60 (m, 1H), 4.68 (m, 1H), 4.81 (t, J = 9.8, 1H), 5.18 (d, J = 3.4, 1H) 5.25 (dd, J ₁ = 2.9, J ₂ = 5.9, 1H), 5.20-5.45 (m, 1H), 6.84 (d, J = 8.3, 2H ), 7.08 (d, J = 8.3, 2H), 7.19-7.28 (m, 1H), 7.34-7.42 (m, 2H), 7.63-7.72 (m, 2H), 7.88 (d, J = 8.3, 2H), 9.00 (s, 1H), 9.57 (m, 1H).

ESI-MC Calculate. for C ₆₁ H ₈₅ N ₇ O ₁₆ : 1171.61. Found: 1172.5 (M + H) ⁺ .

Example 113

Synthesis of N-methylsulfonyl-Pro-OBz (P2)

To a solution of Pro-OBn (HCl) (300 mg, 1.24 mmol) in DCM (25 ml, anhydrous) at 0 ° C. in an Ar atmosphere was added dropwise using a DIPEA syringe (0.7 μl) and methanesulfonyl chloride (116 μl). The reaction mixture was stirred at room temperature overnight. DCM (10 ml) was added and the solution was washed successively with aq. KHSO ₄ (15 ml, 10%), aq. NaHCO ₃ (15 ml, sat.) And brine (15 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give pure P2 (350 mg, 1.24 mmol, quant.) As a white solid. Rf = 0.55 (hex. / AcOEt 1: 1).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.91-2.08 (m, 3H), 2.19-2.31 (m, 1H), 2.93 (s, 3H), 3.38 -3.54 (m, 2H), 3.65 (s, 3H), 4.51 (dd, J ₁ = 3.4, J ₂ = 8.3, 1H), 5.11 (d, J = 12.2, 1H), 5.18 (d, J = 12.2, 1H), 7.31 (m, 5H).

ESI-MC Calculate. for C ₁₃ H ₁₇ NO ₄ S: 283.09. Found: 284.1 (M + H) ⁺ .

Example 114

Synthesis of N-methylsulfonyl-Pro-OH (P1)

Degassed mixture of N-methylsulfonyl-Pro-OBz (P2) (250 mg, 0.88 mmol) and Pd (OH) ₂ / C (20% Pd, 100 mg, 40% m / m) in IPA: H ₂ O ( 26 ml: 13 ml) saturate H ₂ and incubated at 1 atm of hydrogen gas for 3 hours. Then the mixture was filtered through a Teflon filter (0.45 μm) and concentrated in vacuo to give the title compound (128 mg, 75% yield) in as a white solid, without further purification.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.94-2.03 (m, 2H), 2.07-2.14 (m, 1H), 2.22-2.35 (m, 1H) 2.96 (s, 3H), 3.44 (m, 2H), 4.44 (dd, J ₁ = 3.9, J ₂ = 8.8, 2H), 9.10 (m, 1H) .

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 24.96, 31.08, 38.46, 48.06, 60.66, 174.75.

ESI-MC Calculate. for C ₆ H ₁₁ NO ₄ S: 193.22. Found: 194.0 (M + H) ⁺ .

Example 115

Synthesis of [Methylsulfonyl] ⁹ aplidine (9MSAPL1)

Following the procedure developed for the synthesis of 9ZASHPL2, starting from SAPL2 (10 mg, 10.7 μmol) and N-methylsulfonyl-Pro-OH (10 mg, 53 μmol), the title compound (9 mg, 74%) was obtained as white solid after purification by HPLC (Symetry Prep C18, isotrate ratio ACN / H ₂ O 60:40 (flow rate: 3 ml / min, 150 × 7.8 mm, at 270 nm, t _R = 9 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.78-1.00 (m, 24H), 1.20-2.50 (m, 26H), 2.56 (s, 3H), 2.93 (m, 1H), 3.04 (s, 3H), 3.06 (s, 3H), 3.08-3.25 (m, 2H), 3.28-3.50 (m, 2H), 3.60 (m, 2H), 3.70 (m, 1H), 3.79 (s, 3H), 4.05 (m, 2H), 4.17 (m, 1H), 4.60 (m , 2H), 4.81 (m, 2H), 5.10 (m, 1H), 5.19 (d, J = 3.4, 1H), 5.33 (m, 1H), 6.84 ( d, J = 8.3, 2H), 6.86 (m, 1H), 7.07 (d, J = 8.3, 2H), 7.09 (m, 1H), 7.82 (d, J = 9.2, 1H).

ESI-MC Calculate. for C ₅₅ H ₈₇ N ₇ O ₁₅ S: 1117.60. Found: 1118.7 (M + H) ⁺ .

Example 116

Synthesis of [Methylsulfonyl] ⁸ Didemnin A (8MSAPL1)

To a solution of SAPL2 (10 mg, 10.7 μmol) in DCM (200 μl, anhydrous) at 0 ° C. under Ar atmosphere, DIPEA (3 μl) and methanesulfonyl chloride (0.85 μl) were added. The reaction mixture was stirred at 5 ° C. overnight. DCM (10 ml) was added and the solution was washed successively with aq. KHSO ₄ (5 ml, 10%), aq. NaHCO ₃ (5 ml, sat.) And brine (5 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give pure 8MSAPL1 (11 mg, quant.) As a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.83-0.97 (m, 24H), 1.10-1.45 (m, 10H), 1.50-1.65 (m, 5H) 1.76-1.82 (m, 2H), 2.00-2.20 (m, 3H), 2.27-2.36 (m, 1H), 2.45-2.55 (m, 1H), 2.56 (s, 3H), 2.90 (s, 3H), 3.02 (s, 3H), 3.08 (d, J = 16.6, 1H), 3.17 (dd , J ₁ = 10.7, J ₂ = 14.1, 1H), 3.37 (m, 1H), 3.60 (m, 2H), 3.70 (m, 1H), 3.79 (s , 3H), 3.99-4.11 (m, 3H), 4.49 (m, 1H), 4.59 (m, 1H), 4.78 (m, 2H), 5.06 (m, 1H), 5.19 (d, J = 3.9, 1H), 6.68 (d, J = 8.8, 1H), 6.84 (d, J = 8.3, 2H), 7, 07 (d, J = 8.3, 2H), 7.33 (d, J = 9.8, 1H), 7.61 (d, J = 8.8, 1H).

ESI-MC Calculate. for C ₅₀ H ₈₀ N ₆ O ₁₄ S: 1020.55. Found: 1022.1 (M + H) ⁺ .

Example 117

Synthesis of [Biotin] ⁸ Didemnin A (8BISAPL1)

To a solution of HATU (24 mg, 61 μmol), HOAt (8 mg, 63 μmol), SAPL2 (20 mg, 21.4 μmol) and d-biotin (7.8 mg, 32 μmol) in bezv. DCM (400 μl) at 0 ° C. in an Ar atmosphere was added dropwise using an NMM syringe. The resulting mixture was stirred for 2 hours at 0 ° C and then at room temperature for another 14 hours. DCM (10 ml) was added and the solution was washed successively with aq. KHSO ₄ (5 ml, 10%), aq. NaHCO ₃ (5 ml, sat.) And brine (5 ml). The organic phase is dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The title compound (18 mg, 72%) was obtained as a white solid after purification by HPLC (Symetry Prep C18, gradient ACN / H ₂ O 60:40 - 100: 0 in 10 min (flow: 3 ml / min, 150 × 7.8 mm, at 270 nm, t _R = 6 min).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.78-1.00 (m, 24H), 1.20-1.98 (m, 23H), 2.00-2.62 (m, 7H) 2.53 (s, 3H), 2.80-3-00 (m, 2H), 2.86 (s, 3H), 3.16 (m, 2H), 3.35 (m, 2H), 3.57 (m, 2H), 3.70 (m, 1H), 3.79 (s, 3H), 4.01 (m, 2H), 4.10 (m, 1H), 4.31 (m , 1H), 4.46 (m, 1H), 4.56 (m, 1H), 4.82 (m, 2H), 5.00 (m, 1H), 5.14 (m, 2H), 5 67 (s, 1H), 6.23 (s, 1H), 6.84 (d, J = 8.3, 2H), 7.07 (d, J = 8.3, 2H), 7.28 (d, J = 8.5, 1H), 7.35 (d, J = 7.8, 1H), 8.01 (d, J = 8.7, 1H).

ESI-MC Calculate. for C ₅₉ H ₉₂ N ₈ O ₁₄ S: 1168.65. Found: 1169.6 (M + H) ⁺ .

Example 118

Synthesis of [phenylcarbamide] ⁸ didemnin A (8PUSAPL1)

To a solution of SAPL2 (10 mg, 10.7 μmol) in DCM (200 μl, anhydrous) at 0 ° C in an atmosphere of Ar, phenylisocyanate (1.3 μl, 12 μmol) was added and the reaction mixture was stirred at rt. 4 hours. DCM (10 ml) was added and the solution was washed successively with aq. KHSO ₄ (5 ml, 10%), aq. NaHCO ₃ (5 ml, sat.) And brine (5 ml). The organic phase was dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give pure 8PUSAPL1 (11 mg, 94%) as a white solid.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.85-0.94 (m, 24H), 1.11-1.43 (m, 4H), 1.22 (d, J = 6.8, 3H), 1.35 (d, J = 6.8, 3H), 1.49-1.83 (m, 5H), 1.73 (s, 3H), 2.02 (m, 1H), 2 14 (m, 2H), 2.34 (dt, J ₁ = 3.4, J ₂ = 6.8, 1H), 2.54 (s, 3H), 2.91 (s, 3H), 3 04 (d, J = 16.6, 1H), 3.17 (dd, J ₁ = 11.2, J ₂ = 14.6, 1H), 3.36 (dd, J ₁ = 3.9, J ₂ = 14.2, 1H), 3.57 (dd, J ₁ = 4.4, J ₂ = 10.7, 1H), 3.59-3.63 (m, 1H), 3.67- 3.75 (m, 1H), 3.79 (s, 3H), 3.96-4.10 (m, 2H), 4.19 (q, J = 6.8, 1H), 4.58 ( m, 1H), 4.74 (dd, J ₁ = 3.9, J ₂ = 8.8, 1H), 4.78-4.85 (m, 1H), 5.05 (m, 2H), 5.17 (d, J = 3.4, 1H), 6.52 (s, 1H), 6.84 (d, J = 8.3, 2H), 7.03-7.09 (m, 3H ), 7.22-7.32 (m, 4H), 7.39 (d, J = 8.3, 2H), 7.93 (d, J = 8.8, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.70, 15.51, 17.16, 18.74, 21.11, 22.41, 23.29, 23.98, 24.94, 25, 10, 25.32, 26.97, 28.18, 30.36, 31.57, 34.44, 36.51, 38.86, 41.64, 45.23, 47.30, 49.80, 50.08, 54.60, 55.50, 56.06, 57.51, 62.77, 66.45, 68.17, 70.72, 81.93, 114.36, 120.76, 123, 86, 129.12, 130.04, 130.57, 138.78, 141.00, 157.78, 158.87, 169.92, 170.68, 171.49, 172.43, 173.44, 181.47, 192.38, 204.93, 206.66.

ESI-MC Calculate. for C ₅₆ H ₈₃ N ₇ O ₁₃ : 1161.60. Found: 1062.6 (M + H) ⁺ .

Example 119

Synthesis of [phenylthiocarbamide] ⁸ didemnin A (8PTSAPL1)

Following the procedure developed for the synthesis of 8PUSAPL1, starting from SAPL2 (10 mg, 10.7 μmol) and phenylthioisocyanate (1.3 μl, 12 μmol), after 15 hours of stirring, the title compound (11 mg, 93%) is obtained in as a white solid, without further purification.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.84-0.99 (m, 24H), 1.14-1.46 (m, 9H), 1.46-1.78 (m, 9H) 1.98-2.19 (m, 3H), 2.34 (m, 1H), 2.54 (s, 3H), 2.91 (m, 1H), 3.00 (s, 3H), 3.08-3.21 (m, 1H), 3.36 (dd, J ₁ = 4.4, J ₂ = 14.1, 1H), 3.55-3.64 (m, 2H), 3 66-3.74 (m, 1H), 3.79 (s, 3H), 3.96-4.12 (m, 2H), 4.21 (q, J = 6.8, 1H), 4 59 (t, J = 5.3, 1H), 4.75 (dd, J ₁ = 3.4, J ₂ = 8.3, 1H), 4.83 (t, J = 10.3, 1H ), 5.05-5.12 (m, 2H), 5.16 (d, J = 3.4, 1H), 6.30 (t, J = 7.3, 1H), 6.84 (d , J = 8.3, 2H), 7.07 (d, J = 8.3, 2H), 7.21-7.37 (m, 6H), 7.93 (d, J = 8.8, 1H), 8.08 (d, J = 8.8, 1H).

¹³ C-NMR (75 MHz, CDCl ₃ ) δ 11.68, 15.28, 15.85, 17.15, 18.73, 21.12, 22.73, 23.66, 24.01, 25, 13, 25.33, 26.62, 28.16, 29.92, 31.53, 32.42, 32.90, 34.41, 36.77, 38.86, 41.69, 47.28, 50.12, 55.50, 56.48, 57.51, 59.87, 66.47, 70.75, 81.91, 114.37, 125.25, 125.95, 126.57, 127, 54, 129.12, 129.77, 130.06, 130.57, 135.43, 158.88, 168.54, 169.90, 170.70, 171.49, 172.38, 190.41.

ESI-MC Calculate. for C ₅₆ H ₈₃ N ₇ O ₁₂ S: 1077.58. Found: 1078.5 (M + H) ⁺ .

Example 120

Synthesis of [Butylcarbamide] ⁸ Didemnin A (8BUSAPL1)

Following the procedure developed for the synthesis of 8PUSAPL1, starting from SAPL2 (10 mg, 10.7 μmol) and butylthioisocyanate (1.4 μl, 12 μmol), after 4 hours of stirring, the title compound (9 mg, 78%) is obtained in as a white solid, without further purification.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.85-0.94 (m, 24H), 1.10-1.80 (m, 24H), 2.03 (m, 1H), 2.13 (m, 2H), 2.33. (m, 1H), 2.55 (s, 3H), 2.72 (s, 3H), 3.02 (d, J = 16.1, 1H), 3.17 (dd, J ₁ = 11, 2, J ₂ = 14.2, 1H), 3.23-3.40 (m, 3H), 3.62-3.78 (m, 3H), 3.79 (s, 3H), 4.03 (m, 2H), 4.19 (q, J = 6.8, 1H), 4.58 (m, 2H), 4.71 (dd, J ₁ = 3.4, J ₂ = 8.3, 1H), 4.82 (m, 1H), 5.00 (m, 2H), 5.16 (d, J = 3.4, 1H), 5.22-5.28 (m, 2H), 6 85 (d, J = 8.8, 2H), 7.08 (d, J = 8.3, 2H), 7.21-7.29 (m, 1H), 7.96 (d, J = 9.2, 1H).

ESI-MC Calculate. for C ₅₄ H ₈₇ N ₇ O ₁₃ : 1041.64. Found: 1042.7 (M + H) ⁺ .

Example 121

Synthesis of [Butylthiocarbamide] ⁸ Didemnin A (8BTSAPL1)

Following the procedure developed for the synthesis of 8PUSAPL1, starting from SAPL2 (10 mg, 10.7 μmol) and butylthioisocyanate (1.5 μl, 12 μmol), after 15 hours of reaction, the title compound (10 mg, 86%) is obtained in as a white solid, without further purification.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 0.86-0.97 (m, 24H), 1.14-1.78 (m, 27H), 1.99-2.14 (m, 3H) 2.34 (m, 1H), 2.55 (s, 3H), 2.88 (s, 3H), 3.01 (d, J = 16.6, 1H), 3.16 (dd, J ₁ = 11.2, J ₂ = 14.6, 1H), 3.36 (dd, J ₁ = 4.4, J ₂ = 14.2, 1H), 3.49-3.74 (m, 3H ), 3.79 (s, 3H), 4.02 (d, J = 6.8, 2H), 4.21 (q, J = 6.8, 1H), 4.58 (m, 1H ;, 4.70 (dd, J ₁ = 2.9, J ₂ = 8.3, 1H), 4.82 (t, J = 10.3, 1H), 5.07-5.12 (m, 2H) 5.09 (d, J = 3.9, 1H), 5.60 (m, 1H), 6.36 (dd, J ₁ = 5.4, J ₂ = 8.3, 1H), 6, 84 (d, J = 8.8, 2H), 7.08 (d, J = 8.8, 2H), 7.84 (d, J = 8.3, 1H), 7.91 (d, J = 9.3, 1H).

ESI-MC Calculate. for C ₅₄ H ₈₇ N ₇ O ₁₂ S: 1057.61. Found: 1080.7 (M + Na) ⁺ .

Claims (28)

1. The compound of the formula

where X is independently —O— or —NH—; Y is —CO— or —COCHCH ₃ CO—; R ₄ means H or an alkyl group; X ₁ represents O or S; and when Y means —CO— then a) X ₂ independently means CR, O (and R _{2 is} absent) or N, where R independently means H or an organic group selected from an alkyl group, an alkenyl group, an aryl group, an aralkyl group; and R ₁ and R ₂ each independently means H or an organic group selected from an alkyl group, an alkenyl group, an aryl group, an aralkyl group, an amido group RCONH-, where R is an alkyl group, an alkenyl group, an aryl group, an aralkyl group, where when R ₁ and R ₂ each independently represents an organic group selected from an alkyl group, an alkenyl group, an aryl group, an aralkyl group or an amido group RCONH- where R represents an alkenyl group, an aryl group, an aralkyl group, these groups being optionally substituted with het rotsiklicheskoy group which is a heteroaromatic or heteroalicyclic group containing one, two or three heteroatoms selected from nitrogen, oxygen or sulfur; or c) where the aa8 group forms an optionally N-substituted proline of formula

where R ₃ independently means H or an organic group selected from the group RSO ₂ - or an acyl group RCO—, where R is an alkyl group, an alkenyl group, an aryl group, an aralkyl group that are optionally substituted with one or more optionally protected amino groups; or c) R ₁ and R ₂ can form an optionally substituted heterocyclic group with X ₂ that represents a heteroaromatic or heteroalicyclic group containing one, two or three heteroatoms selected from nitrogen, oxygen or sulfur; or d) aa8 is substituted with an organic group selected from the group RSO ₂ -, where R is an alkyl group, an alkenyl group, an aryl group, an aralkyl group; when Y is —COCHCH ₃ CO— then a) X ₂ is N, and R ₁ and R _{2 are} each independently H or an organic group selected from an alkyl group, an alkenyl group, an aryl group, an aralkyl group or an acyl group RCO—, where R is an alkyl group, an alkenyl group, an aryl a group, an aralkyl group, which are optionally substituted with a heterocyclic group which represents a heteroaromatic or heteroalicyclic group containing one, two or three heteroatoms selected from nitrogen, oxygen or sulfur; or C) where the group aa8 forms an optionally N-substituted proline of the formula

where R _{3 is} independently an RSO ₂ - group, where R is an alkyl group that is optionally substituted with one or more optionally protected amino groups, a hydroxy group; c) R ₁ and R ₂ can form a heterocyclic group with X ₂ that represents a heteroaromatic or heteroalicyclic group containing one, two or three heteroatoms selected from nitrogen, oxygen or sulfur; or d) aa8 is substituted with an organic group selected from the group RSO ₂ -, where R is an alkyl group, an alkenyl group, an aryl group, an aralkyl group, with the exception of the compounds of the following formula

and its pharmaceutically acceptable salts. 2. The compound according to claim 1, where X is —NH—. 3. The compound of claim 1, wherein X is —O—. 4. The compound according to any one of claims 1 to 3, where Y is —CO—. 5. The compound according to any one of the preceding paragraphs, where R ₄ means methyl. 6. The compound according to any one of the preceding paragraphs, where X ₁ means O. 7. The compound according to any one of the preceding paragraphs, where Y is CO and X ₂ R ₁ is an optionally substituted aralkyloxy group. 8. The compound according to claim 7, where X ₂ R ₁ means benzyloxy. 9. The compound according to any one of claims 1 to 6, where X ₂ R ₁ means an optionally substituted amino group. 10. The compound of claim 9, wherein X ₂ R ₁ is an —NHR ₁ group, where R ₁ is an optionally substituted alkyl group, alkenyl group, aryl group or an aralkyl group. 11. The compound of claim 10, where R ₁ means an alkyl group or an aryl group. 12. The compound according to claim 11, where R ₁ means a phenyl group or a butyl group. 13. The compound according to any one of claims 1 to 6, where Y is CO and X ₂ R ₁ is an optionally substituted alkyl group. 14. The compound according to item 13, where X ₂ R ₁ means a propyl group, an isopropyl group, a pentyl group or a biotin group. 15. The compound according to any one of claims 1 to 6, where -C (= O) X ₂ R ₁ R ₂ forms an optionally substituted acyl group of an amino acid. 16. The compound of claim 15, wherein the optionally substituted acyl group of the amino acid is an optionally substituted proline. 17. The compound according to clause 16, where the optionally substituted proline means optionally substituted norvalinproline, optionally substituted alanineproline, Boc-proline, optionally substituted alkylproline. 18. The compound according to clause 16, where the optionally substituted proline means norvalineproline, Z-norvalineproline, alanine proline, Z-alanine proline, Boc-alanine proline, isobutyryl proline or optionally protected D-lactylproline. 19. The compound according to any one of claims 1 to 5, where X ₁ means S and X ₂ R ₁ means a group —NHR ₁ , where R ₁ means a substituted alkyl group, an alkenyl group, an aryl group or an aralkyl group. 20. The compound according to item 21, where R ₁ means an alkyl group or an aryl group. 21. The compound according to claim 20, where R ₁ means a phenyl group or a butyl group. 22. The compound according to any one of claims 1 to 6, where R ₁ and R ₂ together with X ₂ form an optionally substituted heterocyclic group. 23. The compound according to item 22, where the heterocyclic group means coumarin. 24. The compound according to any one of claims 1 to 5, where aa8 is substituted by an organic group RSO ₂ -, where R is as defined. 25. The compound of claim 24, wherein R is methyl. 26. The compound according to claim 1, which is selected from the group including 8- [phenylurea] didemnin A, 8- [butylurea] didemnin A, 3- [val] -8- [isobutyryl] didemnin A, 3- [Hiv] -9- [isobutyryl] aplidine, 3- [Val] -9- [isobutyryl] aplidine, 3- [Hiv] -8- [isobutyryl] didemnin A, 3- [Hiv] -9- [Ala] aplidine, 3- [Hiv] -9- [Nva-Pro] aplidine, 8- [phenylthiourea] didemnin A, 8- [coumarin] didemnin A, 8- [butylthiourea] didemnin A, 8- [methylsulfonyl] didemnin A, 3- [val] -Z-didemnin A, 3- [Hiv] -8- [Val] didemnin A, 3- [Hiv] -8- [butyryl] didemnin A, 3- [Hiv] -9- [Z-ala] aplidine, 3- [Hiv] -Z-didemnin A, 3- [Hiv] -9- [Z-Nva-Pro] aplidine, 3- [Hiv] -9- [Boc-Ala] aplidine, 3- [Hiv] -8- [Boc-Val] didemnin A, 3- [Hiv] -8- [Val] -9- [isobutyryl] didemnin A, 3- [Hiv] -8- [hexanoyl] didemnin A, 3- [Hiv] -8- [Pro] didemnin A, 9- [methylsulfonyl] aplidine. 27. The compound of claim 26, which is 8- [phenylurea] didemnin A. 28. The compound according to any one of the preceding paragraphs in the form of a pharmaceutical salt.